Compositions and methods for the treatment of hyperglycemia and metabolic syndrome

ABSTRACT

The invention relates to the compounds and compositions of Formula I, Formula II, Formula III, Formula IV, Formula V, Formula VI or its pharmaceutical acceptable polymorphs, solvates, enantiomers, stereoisomers and hydrates thereof. The pharmaceutical compositions comprises a salt of metformin and the methods for treating or preventing metabolic syndrome, prediabetes and diabetes may be formulated for oral, buccal, rectal, topical, transdermal, transmucosal, intravenous, parenteral administration, syrup, or injection. Such compositions may be used to treatment of diabetes mellitus, obesity, lipid disorders, hypertriglyceridemia, hyperglycemia, hyperinsulinemia and insulin resistance.

PRIORITY

This patent application is a Continuation of, and hereby incorporates byreference the entirety of the disclosures of and claims priority topending U.S. Non-Provisional application Ser. No. 14/938,185 titled‘COMPOSITIONS AND METHODS FOR THE TREATMENT OF HYPERGLYCEMIA ANDMETABOLIC SYNDROME’ filed on Nov. 11, 2015. This further claims thebenefit of PCT Application No. PCT/IB2015/056704 filed on Sep. 3, 2015,which further claims benefit of Indian Provisional Patent ApplicationNo. 1436/CHE/2015 filed on Mar. 21, 2015, the entire disclosure of whichis relied on for all purposes and is incorporated into this applicationby reference.

FIELD OF THE INVENTION

This disclosure generally relates to compounds and compositions for thetreatment of diabetes and pre-diabetes. More particularly, thisinvention relates to treating subjects with a pharmaceuticallyacceptable dose of compounds, crystals, solvates, enantiomer,stereoisomer, hydrates, or mixtures thereof.

BACKGROUND OF THE INVENTION

The multifaceted metabolic syndrome is defined as a number of majormetabolic disorders that enhances the risk of cardiovascular disease(CVD)—still the most important cause of death in the Western world—andtype 2 diabetes mellitus. It is also known as the insulin resistancesyndrome, syndrome X, dysmetabolic syndrome, or the deadly quartet, andis characterized by aberrations in a wide variety of metabolic riskmarkers such as hyperinsulinemia, impaired glucose metabolism, elevatedplasma levels of triglycerides, decreased levels of high-densitylipoprotein cholesterol (HDL-C), raised blood pressure, centrallydistributed obesity, impaired endothelial and haemostatic function, anda low-grade inflammatory state.

Type 2 Diabetes Mellitus (T2DM) is characterized by fasting andpostprandial hyperglycemia and relative insulin insufficiency. If leftuntreated, then hyperglycemia may cause long term microvascular andmacrovascular complications, such as nephropathy, neuropathy,retinopathy, and atherosclerosis. This disease causes significantmorbidity and mortality at considerable expense to patients, theirfamilies and society. The incidence of T2DM worldwide is now increasingat more rapid rates in Africa, Asia and South America than in Europe orthe U.S. Thus, T2DM is now considered worldwide epidemic.

Oxidative stress has long been associated with the late complications ofdiabetes, and has been implicated in their etiology. The reactive oxygenintermediates, produced in mitochondria, peroxisomes, and the cytosol,are scavenged by cellular defending systems, including enzymatic (ex.superoxide dismutase, glutathione peroxidase GPx, glutathione reductaseand catalase) and nonenzymatic antioxidants (ex. glutathione G-SH,thioredoxin, lipoic acid, ubiquinol, albumin, uric acid, flavonoids,vitamins A, C and E, etc.). Some are located in cell membranes, othersin the cytosol, and others in the blood plasma. In diabetes, an alteredoxidative metabolism is a consequence either of the chronic exposure tohyperglycaemia or of the absolute or relative insulin deficit; insulinregulates several reactions involved in oxido-reductive metabolism.Despite strong experimental evidence indicating that oxidative stressmay determine the onset and progression of late-diabetes complicationscontroversy exists about whether the increased oxidative stress ismerely associative rather than causal in diabetes.

Managing acute pathology of often relies on the addressing underlyingpathology and symptoms of the disease. There is currently a need in theart for new compositions to treatment or delay of the onset of diabetesand pre-diabetes and its associated complications progression.

SUMMARY OF THE INVENTION

The present invention provides compounds, compositions containing thesecompounds and methods for using the same to treat, prevent and/orameliorate the effects of the conditions such as diabetes andpre-diabetes.

The invention herein provides compositions comprising of Formula I orpharmaceutical acceptable salts thereof. The invention also providespharmaceutical compositions comprising one or more compounds of FormulaI or intermediates thereof and one or more of pharmaceuticallyacceptable carriers, vehicles or diluents. These compositions may beused in the treatment of diabetes and pre-diabetes and its associatedcomplications.

In certain embodiments, the present invention relates to the compoundsand compositions of Formula I, or pharmaceutically acceptable vehicle,carrier, enantiomer, stereoisomer, polymorph, hydrate, solvate, mixturesor diluent thereof,

with at least one represented by X⁻, or a mixture thereof

Wherein,

X⁻ independently represents

The compositions are typically compounds in the forms of Tri-salt ofbiguanide such as metformin, a biologically active isomer aminoacidL-Leucine in a dipeptide form and an acid moiety represented by X⁻, inwhich the biguanide moiety is protonated and the acid moiety is at leastin partially ionic form. In some instances, however, for exampledepending on the pH of the environment, the composition may be in theform of a mixture of biguanide, L-Leucine in a dipeptide form and anacid component represented by X. The invention also providespharmaceutical compositions comprising compositions of Formula I andpharmaceutically acceptable excipients.

X⁻ in Formula I is independently selected from

eicosapentaenoate, docosapentaenoate, R-lipoate, D-pantothenate,

a molecular conjugate of eicosapentaenoic acid and L-leucine,

a molecular conjugate of docosapentaenoic acid and L-leucine,

a molecular conjugate of eicosapentaenoic acid, fumaric acid andL-leucine,

a molecular conjugate of eicosapentaenoic acid, citric acid andL-leucine,

a molecular conjugate of eicosapentaenoic acid, succinic acid andL-leucine, or

a molecular conjugate of eicosapentaenoic acid and D-pantothenate.

In certain embodiments, the present invention relates to the compoundsand compositions of Formula II, or pharmaceutically acceptable vehicle,carrier, enantiomer, stereoisomer, polymorph, hydrate, solvate, mixturesor diluent thereof,

with at least one represented by X⁻, or a mixture thereof

Wherein,

X⁻ independently represents

The compositions are typically compounds in the forms of Tri-salt ofbiguanide such as metformin, a biologically active isomer aminoacidL-Leucine in a tripeptide form and an acid moiety represented by X⁻, inwhich the biguanide moiety is protonated and the acid moiety is at leastin partially ionic form. In some instances, however, for exampledepending on the pH of the environment, the composition may be in theform of a mixture of biguanide, L-Leucine in a tripeptide form and anacid component represented by X. The invention also providespharmaceutical compositions comprising compositions of Formula II andpharmaceutically acceptable excipients.

X⁻ in Formula II is independently selected from

eicosapentaenoate, docosapentaenoate, R-lipoate, D-pantothenate,

a molecular conjugate of eicosapentaenoic acid and L-leucine,

a molecular conjugate of docosapentaenoic acid and L-leucine,

a molecular conjugate of eicosapentaenoic acid, fumaric acid andL-leucine,

a molecular conjugate of eicosapentaenoic acid, citric acid andL-leucine,

a molecular conjugate of eicosapentaenoic acid, succinic acid andL-leucine, or

a molecular conjugate of eicosapentaenoic acid and D-pantothenate.

In certain embodiments, the present invention relates to the compoundsand compositions of Formula III, or pharmaceutically acceptable vehicle,carrier, enantiomer, stereoisomer, polymorph, hydrate, solvate, mixturesor diluent thereof,

with at least one represented by X⁻, or a mixture thereof

Wherein,

X⁻ independently represents

The compositions are typically compounds in the forms of Tri-salt ofbiguanide such as metformin, a biologically active isomer aminoacidL-Leucine in a tetrapeptide form and an acid moiety represented by X⁻,in which the biguanide moiety is protonated and the acid moiety is atleast in partially ionic form. In some instances, however, for exampledepending on the pH of the environment, the composition may be in theform of a mixture of biguanide, L-Leucine in a tetrapeptide form and anacid component represented by X. The invention also providespharmaceutical compositions comprising compositions of Formula III andpharmaceutically acceptable excipients.

X⁻ in Formula III is independently selected from

eicosapentaenoate, docosapentaenoate, R-lipoate, D-pantothenate,

a molecular conjugate of eicosapentaenoic acid and L-leucine,

a molecular conjugate of docosapentaenoic acid and L-leucine,

a molecular conjugate of eicosapentaenoic acid, fumaric acid andL-leucine,

a molecular conjugate of eicosapentaenoic acid, citric acid andL-leucine,

a molecular conjugate of eicosapentaenoic acid, succinic acid andL-leucine, or

a molecular conjugate of eicosapentaenoic acid and D-pantothenate.

In certain embodiments, the present invention relates to the compoundsand compositions of Formula IV, or pharmaceutically acceptable vehicle,carrier, enantiomer, stereoisomer, polymorph, hydrate, solvate, mixturesor diluent thereof,

Wherein,

R¹ independently represents NULL,

R² independently represents

In certain embodiments, the present invention relates to the compoundsand compositions of Formula V, or pharmaceutically acceptable vehicle,carrier, enantiomer, stereoisomer, polymorph, hydrate, solvate, mixturesor diluent thereof,

Wherein,

R¹ independently represents NULL,

R² independently represents

In certain embodiments, the present invention relates to the compoundsand compositions of Formula VI, or pharmaceutically acceptable vehicle,carrier, enantiomer, stereoisomer, polymorph, hydrate, solvate, mixturesor diluent thereof,

Wherein,

R¹ independently represents NULL,

R² independently represents

The invention further provides methods for treating diabetes (especiallytype 2 diabetes), obesity, cardiac arrhythmia, myocardial infarction andelevated triglycerides. The compounds and compositions of this inventionmay provide high blood levels of the compositions of this invention,when administered to patients, preferably by oral administration.

Herein the application also provides a kit comprising any of thepharmaceutical compositions disclosed herein. The kit may compriseinstructions for use in the treatment of diabetes and pre-diabetes orits related complications.

The application also discloses a pharmaceutical composition comprising apharmaceutically acceptable carrier and any of the compositions herein.In some aspects, the pharmaceutical composition is formulated forsystemic administration, oral administration, sustained release,parenteral administration, injection, subdermal administration, ortransdermal administration.

Herein, the application additionally provides kits comprising thepharmaceutical compositions described herein. The kits may furthercomprise instructions for use in the treatment of diabetes andpre-diabetes or its related complications.

The compositions described herein have several uses. The presentapplication provides, for example, methods of treating a patientsuffering from diabetes and pre-diabetes or its related complicationsmanifested from metabolic or genetic conditions or disorders, metabolicdiseases, chronic diseases or disorders; neurodegenerative disorders,metabolic condition, Hepatology, Cancer, Respiratory, Hematological,Orthopedic, Cardiovascular, Renal, Skin, Vascular or Ocularcomplications.

In the illustrative embodiments, examples of compounds of Formula I andFormula IV are as set forth below:

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1: compound 1 of Example1, 1H NMR (DMSO-d6) data.

FIG. 2: compound 5 of Example2, 1H NMR (DMSO-d6) data.

DETAILED DESCRIPTION OF THE INVENTION Definitions

As used herein, the following terms and phrases shall have the meaningsset forth below. Unless defined otherwise, all technical and scientificterms used herein have the same meaning as commonly understood to one ofordinary skill in the art.

The compounds of the present invention can be present in the form ofpharmaceutically acceptable salts. The compounds of the presentinvention can also be present in the form of pharmaceutically acceptableesters (i.e., the methyl and ethyl esters of the acids of Formula I,Formula II, Formula III, Formula IV, Formula V or Formula VI to be usedas prodrugs). The compounds of the present invention can also besolvated, i.e. hydrated. The solvation can be affected in the course ofthe manufacturing process or can take place i.e. as a consequence ofhygroscopic properties of an initially anhydrous compound of Formula I,Formula II, Formula III, Formula IV, Formula V or Formula VI(hydration).

Compounds that have the same molecular Formula but differ in the natureor sequence of bonding of their atoms or the arrangement of their atomsin space are termed “isomers.” Isomers that differ in the arrangement oftheir atoms in space are termed “stereoisomers.” Diastereomers arestereoisomers with opposite configuration at one or more chiral centerswhich are not enantiomers. Stereoisomers bearing one or more asymmetriccenters that are non-superimposable mirror images of each other aretermed “enantiomers.” When a compound has an asymmetric center, forexample, if a carbon atom is bonded to four different groups, a pair ofenantiomers is possible. An enantiomer can be characterized by theabsolute configuration of its asymmetric center or centers and isdescribed by the R- and S-sequencing rules of Cahn, Ingold and Prelog,or by the manner in which the molecule rotates the plane of polarizedlight and designated as dextrorotatory or levorotatory (i.e., as (+) or(−)-isomers respectively). A chiral compound can exist as eitherindividual enantiomer or as a mixture thereof. A mixture containingequal proportions of the enantiomers is called a “racemic mixture”.

Compounds and compositions of Formula I, Formula II, Formula III,Formula IV, Formula V and Formula VI contains biologically active formof L-isomer of Leucine in either dipeptide, tripeptide or tetrapeptideform individually or in a molecular conjugate form with apolyunsaturated fatty acid.

As used herein, the term “metabolic condition” refers to an Inbornerrors of metabolism (or genetic metabolic conditions) are geneticdisorders that result from a defect in one or more metabolic pathways;specifically, the function of an enzyme is affected and is eitherdeficient or completely absent. Metabolic or genetic disorders orcondition associated diseases include: Hepatic, Neurologic, Psychiatric,Hematologic, Respiratory, Renal, Cardiovascular, Cancer,Musculoskeletal, Orthopedic and Gastrointestinal.

Metabolic syndrome related disorders includes such as chronic diabetes,type 2 diabetes, cancer cachexia, type 1 diabetes, obesity, fatty liverdisease, NASH, NAFLD, Pre-diabetes, lipid disorders, hyperinsulinemia,insulin resistance and other related diseases or any other medicalcondition, is well understood in the art, and includes administration ofa composition which reduces the frequency of, or delays the onset of,symptoms of a medical condition in a subject relative to a subject whichdoes not receive the composition.

The term “polymorph” as used herein is art-recognized and refers to onecrystal structure of a given compound.

The phrases “parenteral administration” and “administered parenterally”as used herein refer to modes of administration other than enteral andtopical administration, such as injections, and include withoutlimitation intravenous, intramuscular, intrapleural, intravascular,intrapericardial, intraarterial, intrathecal, intracapsular,intraorbital, intracardiac, intradennal, intraperitoneal, transtracheal,subcutaneous, subcuticular, intra-articular, subcapsular, subarachnoid,intraspinal and intrastemal injection and infusion.

A “patient,” “subject,” or “host” to be treated by the subject methodmay mean either a human or non-human animal, such as primates, mammals,and vertebrates.

The phrase “pharmaceutically acceptable” is art-recognized. In certainembodiments, the term includes compositions, polymers and othermaterials and/or dosage forms which are, within the scope of soundmedical judgment, suitable for use in contact with the tissues ofmammals, human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

The phrase “pharmaceutically acceptable carrier” is art-recognized, andincludes, for example, pharmaceutically acceptable materials,compositions or vehicles, such as a liquid or solid filler, diluent,solvent or encapsulating material involved in carrying or transportingany subject composition, from one organ, or portion of the body, toanother organ, or portion of the body. Each carrier must be “acceptable”in the sense of being compatible with the other ingredients of a subjectcomposition and not injurious to the patient. In certain embodiments, apharmaceutically acceptable carrier is non-pyrogenic. Some examples ofmaterials which may serve as pharmaceutically acceptable carriersinclude: (1) sugars, such as lactose, glucose and sucrose; (2) starches,such as corn starch and potato starch; (3) cellulose, and itsderivatives, such as sodium carboxymethyl cellulose, ethyl cellulose andcellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7)talc; (8) cocoa butter and suppository waxes; (9) oils, such as peanutoil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil andsoybean oil; (10) glycols, such as propylene glycol; (11) polyols, suchas glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters,such as ethyl oleate and ethyl laurate; (13) agar; (14) bufferingagents, such as magnesium hydroxide and aluminum hydroxide; (15) alginicacid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer'ssolution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21)other non-toxic compatible substances employed in pharmaceuticalformulations.

The term “polymorph” as used herein is art-recognized and refers to onecrystal structure of a given compound.

The term “prodrug” is intended to encompass compounds that, underphysiological conditions, are converted into the therapeutically activeagents of the present invention. A common method for making a prodrug isto include selected moieties that are hydrolyzed under physiologicalconditions to reveal the desired molecule. In other embodiments, theprodrug is converted by an enzymatic activity of the host animal.

The term “prophylactic or therapeutic” treatment is art-recognized andincludes administration to the host of one or more of the subjectcompositions. If it is administered prior to clinical manifestation ofthe unwanted condition (e.g., disease or other unwanted state of thehost animal) then the treatment is prophylactic, i.e., it protects thehost against developing the unwanted condition, whereas if it isadministered after manifestation of the unwanted condition, thetreatment is therapeutic, (i.e., it is intended to diminish, ameliorate,or stabilize the existing unwanted condition or side effects thereof).

The term “predicting” as used herein refers to assessing the probabilityaccording to which a condition or disorder such as diabetes andpre-diabetes or related diseases patient will suffer from abnormalitiesor complication and/or death (i.e. mortality) within a defined timewindow (predictive window) in the future. The mortality may be caused bythe central nervous system or complication. The predictive window is aninterval in which the subject will develop one or more of the saidcomplications according to the predicted probability. The predictivewindow may be the entire remaining lifespan of the subject upon analysisby the method of the present invention. Preferably, however, thepredictive window is an interval of one month, six months or one, two,three, four, five or ten years after appearance of the inflammatorycomplication (more preferably and precisely, after the sample to beanalyzed by the method of the present invention has been obtained). Aswill be understood by those skilled in the art, such an assessment isusually not intended to be correct for 100% of the subjects to beanalyzed. The term, however, requires that the assessment will be validfor a statistically significant portion of the subjects to be analyzed.Whether a portion is statistically significant can be determined withoutfurther ado by the person skilled in the art using various well knownstatistic evaluation tools, e.g., determination of confidence intervals,p-value determination, Student's t-test, Mann-Whitney test, etc. Detailsare found in Dowdy and Wearden, Statistics for Research, John Wiley &Sons, New York 1983. Preferred confidence intervals are at least 90%, atleast 95%, at least 97%, at least 98% or at least 99%. The p-values are,preferably, 0.1, 0.05, 0.01, 0.005, or 0.0001. Preferably, theprobability envisaged by the present invention allows that theprediction will be correct for at least 60%, at least 70%, at least 80%,or at least 90% of the subjects of a given cohort.

The term “treating” is art-recognized and includes preventing a disease,disorder or condition from occurring in an animal which may bepredisposed to the disease, disorder and/or condition but has not yetbeen diagnosed as having it; inhibiting the disease, disorder orcondition, e.g., impeding its progress; and relieving the disease,disorder, or condition, e.g., causing regression of the disease,disorder and/or condition. Treating the disease or condition includesameliorating at least one symptom of the particular disease orcondition, even if the underlying pathophysiology is not affected, suchas treating the diabetes and pre-diabetes or condition or disorders suchas diabetes and pre-diabetes condition of a subject by administration ofan agent even though such agent does not treat the cause of thecondition. The term “treating”, “treat” or “treatment” as used hereinincludes curative, preventative (e.g., prophylactic), adjunct andpalliative treatment.

The phrase “therapeutically effective amount” is an art-recognized term.In certain embodiments, the term refers to an amount of a salt orcomposition disclosed herein that produces some desired effect at areasonable benefit/risk ratio applicable to any medical treatment. Incertain embodiments, the term refers to that amount necessary orsufficient to eliminate or reduce medical symptoms for a period of time.The effective amount may vary depending on such factors as the diseaseor condition being treated, the particular targeted constructs beingadministered, the size of the subject, or the severity of the disease orcondition. One of ordinary skill in the art may empirically determinethe effective amount of a particular composition without necessitatingundue experimentation.

In certain embodiments, the pharmaceutical compositions described hereinare formulated in a manner such that said compositions will be deliveredto a patient in a therapeutically effective amount, as part of aprophylactic or therapeutic treatment. The desired amount of thecomposition to be administered to a patient will depend on absorption,inactivation, and excretion rates of the drug as well as the deliveryrate of the salts and compositions from the subject compositions. It isto be noted that dosage values may also vary with the severity of thecondition to be alleviated. It is to be further understood that for anyparticular subject, specific dosage regimens should be adjusted overtime according to the individual need and the professional judgment ofthe person administering or supervising the administration of thecompositions. Typically, dosing will be determined using techniquesknown to one skilled in the art.

Additionally, the optimal concentration and/or quantities or amounts ofany particular salt or composition may be adjusted to accommodatevariations in the treatment parameters. Such treatment parametersinclude the clinical use to which the preparation is put, e.g., the sitetreated, the type of patient, e.g., human or non-human, adult or child,and the nature of the disease or condition.

In certain embodiments, the dosage of the subject compositions providedherein may be determined by reference to the plasma concentrations ofthe therapeutic composition or other encapsulated materials. Forexample, the maximum plasma concentration (Cmax) and the area under theplasma concentration-time curve from time 0 to infinity may be used.

The term “solvate” as used herein, refers to a compound formed bysolvation (e.g., a compound formed by the combination of solventmolecules with molecules or ions of the solute).

When used with respect to a pharmaceutical composition or othermaterial, the term “sustained release” is art-recognized. For example, asubject composition which releases a substance over time may exhibitsustained release characteristics, in contrast to a bolus typeadministration in which the entire amount of the substance is madebiologically available at one time. For example, in particularembodiments, upon contact with body fluids including blood, spinalfluid, mucus secretions, lymph or the like, one or more of thepharmaceutically acceptable excipients may undergo gradual or delayeddegradation (e.g., through hydrolysis) with concomitant release of anymaterial incorporated therein, e.g., an therapeutic and/or biologicallyactive salt and/or composition, for a sustained or extended period (ascompared to the release from a bolus). This release may result inprolonged delivery of therapeutically effective amounts of any of thetherapeutic agents disclosed herein.

The phrases “systemic administration,” “administered systemically,”“peripheral administration” and “administered peripherally” areart-recognized, and include the administration of a subject composition,therapeutic or other material at a site remote from the disease beingtreated. Administration of an agent for the disease being treated, evenif the agent is subsequently distributed systemically, may be termed“local” or “topical” or “regional” administration, other than directlyinto the central nervous system, e.g., by subcutaneous administration,such that it enters the patient's system and, thus, is subject tometabolism and other like processes.

The phrase “therapeutically effective amount” is an art-recognized term.In certain embodiments, the term refers to an amount of a salt orcomposition disclosed herein that produces some desired effect at areasonable benefit/risk ratio applicable to any medical treatment. Incertain embodiments, the term refers to that amount necessary orsufficient to eliminate or reduce medical symptoms for a period of time.The effective amount may vary depending on such factors as the diseaseor condition being treated, the particular targeted constructs beingadministered, the size of the subject, or the severity of the disease orcondition. One of ordinary skill in the art may empirically determinethe effective amount of a particular composition without necessitatingundue experimentation.

The present disclosure also contemplates prodrugs of the compositionsdisclosed herein, as well as pharmaceutically acceptable salts of saidprodrugs.

This application also discloses a pharmaceutical composition comprisinga pharmaceutically acceptable carrier and the composition of a compoundof Formula I, Formula II, Formula III, Formula IV, Formula V or FormulaVI may be formulated for systemic or topical or oral administration. Thepharmaceutical composition may be also formulated for oraladministration, oral solution, injection, subdermal administration, ortransdermal administration. The pharmaceutical composition may furthercomprise at least one of a pharmaceutically acceptable stabilizer,diluent, surfactant, filler, binder, and lubricant.

In many embodiments, the pharmaceutical compositions described hereinwill incorporate the disclosed compounds and compositions (Formula I,Formula II, Formula III, Formula IV, Formula V or Formula VI) to bedelivered in an amount sufficient to deliver to a patient atherapeutically effective amount of a compound of Formula I, Formula II,Formula III, Formula IV, Formula V or Formula VI or composition as partof a prophylactic or therapeutic treatment. The desired concentration ofFormula I, Formula II, Formula III, Formula IV, Formula V or Formula VIor its pharmaceutical acceptable salts will depend on absorption,inactivation, and excretion rates of the drug as well as the deliveryrate of the salts and compositions from the subject compositions. It isto be noted that dosage values may also vary with the severity of thecondition to be alleviated. It is to be further understood that for anyparticular subject, specific dosage regimens should be adjusted overtime according to the individual need and the professional judgment ofthe person administering or supervising the administration of thecompositions. Typically, dosing will be determined using techniquesknown to one skilled in the art.

Additionally, the optimal concentration and/or quantities or amounts ofany particular compound of Formula I, Formula II, Formula III, FormulaIV, Formula V or Formula VI may be adjusted to accommodate variations inthe treatment parameters. Such treatment parameters include the clinicaluse to which the preparation is put, e.g., the site treated, the type ofpatient, e.g., human or non-human, adult or child, and the nature of thedisease or condition.

The concentration and/or amount of any compound of Formula I, FormulaII, Formula III, Formula IV, Formula V or Formula VI may be readilyidentified by routine screening in animals, e.g., rats, by screening arange of concentration and/or amounts of the material in question usingappropriate assays. Known methods are also available to assay localtissue concentrations, diffusion rates of the salts or compositions, andlocal blood flow before and after administration of therapeuticformulations disclosed herein. One such method is microdialysis, asreviewed by T. E. Robinson et al., 1991, microdialysis in theneurosciences, Techniques, volume 7, Chapter 1. The methods reviewed byRobinson may be applied, in brief, as follows. A microdialysis loop isplaced in situ in a test animal. Dialysis fluid is pumped through theloop. When compounds with Formula I, Formula II, Formula III, FormulaIV, Formula V or Formula VI such as those disclosed herein are injectedadjacent to the loop, released drugs are collected in the dialysate inproportion to their local tissue concentrations. The progress ofdiffusion of the salts or compositions may be determined thereby withsuitable calibration procedures using known concentrations of salts orcompositions.

In certain embodiments, the dosage of the subject compounds of FormulaI, Formula II, Formula III, Formula IV, Formula V or Formula VI providedherein may be determined by reference to the plasma concentrations ofthe therapeutic composition or other encapsulated materials. Forexample, the maximum plasma concentration (Cmax) and the area under theplasma concentration-time curve from time 0 to infinity may be used.

Generally, in carrying out the methods detailed in this application, aneffective dosage for the compounds of Formula I, Formula II, FormulaIII, Formula IV, Formula V or Formula VI is in the range of about 0.01mg/kg/day to about 100 mg/kg/day in single or divided. The compounds ofFormula I, Formula II, Formula III, Formula IV, Formula V or Formula VImay be administered at a dose of, for example, less than 0.2 mg/kg/day,0.5 mg/kg/day, 1.0 mg/kg/day, 5 mg/kg/day, 10 mg/kg/day, 20 mg/kg/day,30 mg/kg/day, or 40 mg/kg/day. Compounds of Formula I, Formula II,Formula III, Formula IV, Formula V or Formula VI may also beadministered to a human patient at a dose of, for example, between 0.1mg and 1000 mg, between 5 mg and 80 mg, or less than 1.0, 9.0, 12.0,20.0, 50.0, 75.0, 100, 300, 400, 500, 800, 1000 mg per day. In certainembodiments, the compositions herein are administered at an amount thatis less than 95%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, or 10% of thecompound of Formula I, Formula II, Formula III, Formula IV, Formula V orFormula VI required for the same therapeutic benefit.

An effective amount of the compounds of Formula I, Formula II, FormulaIII, Formula IV, Formula V or Formula VI described herein refers to theamount of one of said salts or compositions which is capable ofinhibiting or preventing a disease. For example diabetes andpre-diabetes or any other metabolic condition or metabolic disorder orany other medical condition.

An effective amount may be sufficient to prohibit, treat, alleviate,ameliorate, halt, restrain, slow or reverse the progression, or reducethe severity of a complication resulting from inflammatory diseasesand/or elevated reactive oxidative-nitrosative species and/orabnormalities in homeostasis's, in patients who are at risk for suchcomplications. As such, these methods include both medical therapeutic(acute) and/or prophylactic (prevention) administration as appropriate.The amount and timing of compositions administered will, of course, bedependent on the subject being treated, on the severity of theaffliction, on the manner of administration and on the judgment of theprescribing physician. Thus, because of patient-to-patient variability,the dosages given above are a guideline and the physician may titratedoses of the drug to achieve the treatment that the physician considersappropriate for the patient. In considering the degree of treatmentdesired, the physician must balance a variety of factors such as age ofthe patient, presence of preexisting disease, as well as presence ofother diseases.

The compositions provided by this application may be administered to asubject in need of treatment by a variety of conventional routes ofadministration, including orally, topically, parenterally, e.g.,intravenously, subcutaneously or intramedullary. Further, thecompositions may be administered intranasally, as a rectal suppository,or using a “flash” formulation, i.e., allowing the medication todissolve in the mouth without the need to use water. Furthermore, thecompositions may be administered to a subject in need of treatment bycontrolled release dosage forms, site specific drug delivery,transdermal drug delivery, patch (active/passive) mediated drugdelivery, by stereotactic injection, or in nanoparticles.

The compositions may be administered alone or in combination withpharmaceutically acceptable carriers, vehicles or diluents, in eithersingle or multiple doses. Suitable pharmaceutical carriers, vehicles anddiluents include inert solid diluents or fillers, sterile aqueoussolutions and various organic solvents. The pharmaceutical compositionsformed by combining the compositions and the pharmaceutically acceptablecarriers, vehicles or diluents are then readily administered in avariety of dosage forms such as tablets, powders, lozenges, syrups,injectable solutions and the like. These pharmaceutical compositionscan, if desired, contain additional ingredients such as flavorings,binders, excipients and the like. Thus, for purposes of oraladministration, tablets containing various excipients such asL-arginine, sodium citrate, calcium carbonate and calcium phosphate maybe employed along with various disintegrates such as starch, alginicacid and certain complex silicates, together with binding agents such aspolyvinylpyrrolidone, sucrose, gelatin and acacia. Additionally,lubricating agents such as magnesium stearate, sodium lauryl sulfate andtalc are often useful for tabletting purposes. Solid compositions of asimilar type may also be employed as fillers in soft and hard filledgelatin capsules. Appropriate materials for this include lactose or milksugar and high molecular weight polyethylene glycols. When aqueoussuspensions or elixirs are desired for oral administration, theessential active ingredient therein may be combined with varioussweetening or flavoring agents, coloring matter or dyes and, if desired,emulsifying or suspending agents, together with diluents such as water,ethanol, propylene glycol, glycerin and combinations thereof. Thecompounds of Formula I, Formula II, Formula III, Formula IV, Formula Vor Formula VI may also comprise enterically coated comprising of variousexcipients, as is well known in the pharmaceutical art.

For parenteral administration, solutions of the compositions may beprepared in (for example) sesame or peanut oil, aqueous propyleneglycol, or in sterile aqueous solutions may be employed. Such aqueoussolutions should be suitably buffered if necessary and the liquiddiluent first rendered isotonic with sufficient saline or glucose. Theseparticular aqueous solutions are especially suitable for intravenous,intramuscular, subcutaneous and intraperitoneal administration. In thisconnection, the sterile aqueous media employed are all readily availableby standard techniques known to those skilled in the art.

The formulations, for instance tablets, may contain e.g. 10 to 100, 50to 250, 150 to 500 mg, or 350 to 800 mg e.g. 10, 50, 100, 300, 500, 700,800 mg of the compounds of Formula I, Formula II, Formula III, FormulaIV, Formula V or Formula VI disclosed herein, for instance, compounds ofFormula I, Formula II, Formula III, Formula IV, Formula V or Formula VIor pharmaceutical acceptable salts of a compounds of Formula I, FormulaII, Formula III, Formula IV, Formula V or Formula VI.

Generally, a composition as described herein may be administered orally,or parenterally (e.g., intravenous, intramuscular, subcutaneous orintramedullary). Topical administration may also be indicated, forexample, where the patient is suffering from gastrointestinal disorderthat prevent oral administration, or whenever the medication is bestapplied to the surface of a tissue or organ as determined by theattending physician. Localized administration may also be indicated, forexample, when a high dose is desired at the target tissue or organ. Forbuccal administration the active composition may take the form oftablets or lozenges formulated in a conventional manner.

The dosage administered will be dependent upon the identity of thediabetes and pre-diabetes; the type of host involved, including its age,health and weight; the kind of concurrent treatment, if any; thefrequency of treatment and therapeutic ratio.

Illustratively, dosage levels of the administered active ingredientsare: intravenous, 0.1 to about 200 mg/kg; intramuscular, 1 to about 500mg/kg; orally, 5 to about 1000 mg/kg; intranasal instillation, 5 toabout 1000 mg/kg; and aerosol, 5 to about 1000 mg/kg of host bodyweight.

Expressed in terms of concentration, an active ingredient can be presentin the compositions of the present invention for localized use about thecutis, intranasally, pharyngolaryngeally, bronchially, intravaginally,rectally, or ocularly in a concentration of from about 0.01 to about 50%w/w of the composition; preferably about 1 to about 20% w/w of thecomposition; and for parenteral use in a concentration of from about0.05 to about 50% w/v of the composition and preferably from about 5 toabout 20% w/v.

The compositions of the present invention are preferably presented foradministration to humans and animals in unit dosage forms, such astablets, capsules, pills, powders, granules, suppositories, sterileparenteral solutions or suspensions, sterile non-parenteral solutions ofsuspensions, and oral solutions or suspensions and the like, containingsuitable quantities of an active ingredient. For oral administrationeither solid or fluid unit dosage forms can be prepared.

Powders are prepared quite simply by comminuting the active ingredientto a suitably fine size and mixing with a similarly comminuted diluent.The diluent can be an edible carbohydrate material such as lactose orstarch. Advantageously, a sweetening agent or sugar is present as wellas flavoring oil.

Capsules are produced by preparing a powder mixture as hereinbeforedescribed and filling into formed gelatin sheaths. Advantageously, as anadjuvant to the filling operation, a lubricant such as talc, magnesiumstearate, calcium stearate and the like is added to the powder mixturebefore the filling operation.

Soft gelatin capsules are prepared by machine encapsulation of slurry ofactive ingredients with an acceptable vegetable oil, light liquidpetrolatum or other inert oil or triglyceride.

Tablets are made by preparing a powder mixture, granulating or slugging,adding a lubricant and pressing into tablets. The powder mixture isprepared by mixing an active ingredient, suitably comminuted, with adiluent or base such as starch, lactose, kaolin, dicalcium phosphate andthe like. The powder mixture can be granulated by wetting with a bindersuch as corn syrup, gelatin solution, methylcellulose solution or acaciamucilage and forcing through a screen. As an alternative to granulating,the powder mixture can be slugged, i.e., ran through the tablet machineand the resulting imperfectly formed tablets broken into pieces (slugs).The slugs can be lubricated to prevent sticking to the tablet-formingdies by means of the addition of stearic acid, a stearic salt, talc ormineral oil. The lubricated mixture is then compressed into tablets.

Advantageously, the tablet can be provided with a protective coatingconsisting of a sealing coat or enteric coat of shellac, a coating ofsugar and methylcellulose and polish coating of carnauba wax.

Fluid unit dosage forms for oral administration such as in syrups,elixirs and suspensions can be prepared wherein each teaspoonful ofcomposition contains a predetermined amount of an active ingredient foradministration. The water-soluble forms can be dissolved in an aqueousvehicle together with sugar, flavoring agents and preservatives to formsyrup. An elixir is prepared by using a hydroalcoholic vehicle withsuitable sweeteners together with a flavoring agent. Suspensions can beprepared of the insoluble forms with a suitable vehicle with the aid ofa suspending agent such as acacia, tragacanth, methylcellulose and thelike.

For parenteral administration, fluid unit dosage forms are preparedutilizing an active ingredient and a sterile vehicle, water beingpreferred. The active ingredient, depending on the form andconcentration used, can be either suspended or dissolved in the vehicle.In preparing solutions the water-soluble active ingredient can bedissolved in water for injection and filter sterilized before fillinginto a suitable vial or ampule and sealing. Advantageously, adjuvantssuch as a local anesthetic, preservative and buffering agents can bedissolved in the vehicle. Parenteral suspensions are prepared insubstantially the same manner except that an active, ingredient issuspended in the vehicle instead of being dissolved and sterilizationcannot be accomplished by filtration. The active ingredient can besterilized by exposure to ethylene oxide before suspending in thesterile vehicle. Advantageously, a surfactant or wetting agent isincluded in the composition to facilitate uniform distribution of theactive ingredient.

In addition to oral and parenteral administration, the rectal andvaginal routes can be utilized. An active ingredient can be administeredby means of a suppository. A vehicle which has a melting point at aboutbody temperature or one that is readily soluble can be utilized. Forexample, cocoa butter and various polyethylene glycols (Carbowaxes) canserve as the vehicle.

For intranasal instillation, a fluid unit dosage form is preparedutilizing an active ingredient and a suitable pharmaceutical vehicle,preferably P.F. water, a dry powder can be formulated when insufflationis the administration of choice.

For use as aerosols, the active ingredients can be packaged in apressurized aerosol container together with a gaseous or liquifiedpropellant, for example, dichlorodifluoromethane, carbon dioxide,nitrogen, propane, and the like, with the usual adjuvants such ascosolvents and wetting agents, as may be necessary or desirable.

The term “unit dosage form” as used in the specification and claimsrefers to physically discrete units suitable as unitary dosages forhuman and animal subjects, each unit containing a predetermined quantityof active material calculated to produce the desired therapeutic effectin association with the required pharmaceutical diluent, carrier orvehicle. The specifications for the novel unit dosage forms of thisinvention are dictated by and are directly dependent on (a) the uniquecharacteristics of the active material and the particular therapeuticeffect to be achieved, and (b) the limitation inherent in the art ofcompounding such an active material for therapeutic use in humans, asdisclosed in this specification, these being features of the presentinvention. Examples of suitable unit dosage forms in accord with thisinvention are tablets, capsules, troches, suppositories, powder packets,wafers, cachets, teaspoonfuls, tablespoonfuls, dropperfuls, ampules,vials, segregated multiples of any of the foregoing, and other forms asherein described.

The tablets of the present invention contain one or morepharmaceutically active agents that are released therefrom upon contactof the tablet with a liquid medium, for example a dissolution mediumsuch as gastrointestinal fluids. “Water soluble,” as used herein inconnection with non-polymeric materials, shall mean from sparinglysoluble to very soluble, i.e., not more than 100 parts water required todissolve 1 part of the non-polymeric, water soluble solute. SeeRemington, The Science and Practice of Pharmacy, pp 208-209 (2000).“Water soluble,” as used herein in connection with polymeric materials,shall mean that the polymer swells in water and can be dispersed at themolecular level or dissolved in water.

As used herein, the term “modified release” shall apply to tablets,matrices, particles, coatings, portions thereof, or compositions thatalter the release of an pharmaceutically active agent in any manner.Types of modified release include controlled, prolonged, sustained,extended, delayed, pulsatile, repeat action, and the like. Suitablemechanisms for achieving these types of modified release includediffusion, erosion, surface area control via geometry and/or impermeablebarriers, or other mechanisms known in the art.

In one embodiment of the invention, the first pharmaceutically activeagent and the hydrophilic polymer are mixed with a powder containing apharmaceutically-acceptable carrier, which is also defined herein as thetablet matrix. In one embodiment, the powder has an average particlesize of about 50 microns to about 500 microns, such as between 50microns and 300 microns. Particles in this size range are particularlyuseful for direct compression processes. In embodiment, the componentsof powder are blended together, for example as dry powders, and fed intothe die cavity of an apparatus that applies pressure to form a tabletcore. Any suitable compacting apparatus may be used, including, but notlimited to, conventional unitary or rotary tablet press. In oneembodiment, the tablet core may be formed by compaction using a rotarytablet press (e.g., such as those commercially available from FetteAmerica Inc., Rockaway, N.J., or Manesty Machines LTD, Liverpool, UK).In general, a metered volume of powder is filled into a die cavity(where the powder is either gravity fed or mechanically fed from afeeder) of the rotary tablet press, and the cavity rotates as part of a“die table” from the filling position to a compaction position. At thecompaction position, the powder is compacted between an upper and alower punch, then the resulting tablet core is pushed from the diecavity by the lower punch and then guided to an injection chute by astationary “take-off bar.

In one embodiment of the invention, the tablet core may be a directlycompressed tablet core made from a powder that is substantially free ofwater-soluble polymeric binders and hydrated polymers. As used herein,what is meant by “substantially free” is less than 5 percent, such asless than 1 percent, such as less than 0.1 percent, such as completelyfree (e.g., 0 percent). This composition is advantageous for minimizingprocessing and material costs and providing for optimal physical andchemical stability of the tablet core. In one embodiment, the density ofthe tablet core is greater than about 0.9 g/cc.

The tablet core may have one of a variety of different shapes. Forexample, the tablet core may be shaped as a polyhedron, such as a cube,pyramid, prism, or the like; or may have the geometry of a space figurewith some non-flat faces, such as a cone, truncated cone, cylinder,sphere, torus, or the like. In certain embodiments, a tablet core hasone or more major faces. For example, the tablet core surface typicallyhas opposing upper and lower faces formed by contact with the upper andlower punch faces in the compression machine. In such embodiments thetablet core surface typically further includes a “belly-band” locatedbetween the upper and lower faces, and formed by contact with the diewalls in the compression machine.

As discussed above, the tablet core contains one or more hydrophilicpolymers. Suitable hydrophilic polymers include, but are not limited to,water swellable cellulose derivatives, polyalkylene glycols,thermoplastic polyalkylene oxides, acrylic polymers, hydrocolloids,clays, gelling starches, swelling cross-linked polymers, and mixturesthereof. Examples of suitable water swellable cellulose derivativesinclude, but are not limited to, sodium carboxymethylcellulose,cross-linked hydroxypropylcellulose, hydroxypropyl cellulose (HPC),hydroxypropylmethylcellulose (HPMC), hydroxyisopropylcellulose,hydroxybutylcellulose, hydroxyphenylcellulose, hydroxyethylcellulose(HEC), hydroxypentylcellulose, hydroxypropylethylcellulose,hydroxypropylbutylcellulose, and hydroxypropylethylcellulose, andmixtures thereof. Examples of suitable polyalkylene glycols include, butare not limited to, polyethylene glycol. Examples of suitablethermoplastic polyalkylene oxides include, but are not limited to,poly(ethylene oxide). Examples of suitable acrylic polymers include, butare not limited to, potassium methacrylatedivinylbenzene copolymer,polymethylmethacrylate, high-molecular weight crosslinked acrylic acidhomopolymers and copolymers such as those commercially available fromNoveon Chemicals under the tradename CARBOPOL™. Examples of suitablehydrocolloids include, but are not limited to, alginates, agar, guargum, locust bean gum, kappa carrageenan, iota carrageenan, tara, gumarabic, tragacanth, pectin, xanthan gum, gellan gum, maltodextrin,galactomannan, pusstulan, laminarin, scleroglucan, gum arabic, inulin,pectin, gelatin, whelan, rhamsan, zooglan, methylan, chitin,cyclodextrin, chitosan, and mixtures thereof. Examples of suitable claysinclude, but are not limited to, smectites such as bentonite, kaolin,and laponite; magnesium trisilicate; magnesium aluminum silicate; andmixtures thereof. Examples of suitable gelling starches include, but arenot limited to, acid hydrolyzed starches, swelling starches such assodium starch glycolate and derivatives thereof, and mixtures thereof.Examples of suitable swelling cross-linked polymers include, but are notlimited to, cross-linked polyvinyl pyrrolidone, cross-linked agar, andcross-linked carboxymethylcellulose sodium, and mixtures thereof.

In one embodiment, an osmogen is incorporated into the tablet core inorder to draw water into the tablet upon contact with fluids, such asgastrointestinal fluids. An osmogen as used herein is a water solublecomponent which preferentially draws water into the tablet core for thepurposes of distributing the water throughout the core, so that theactive ingredient contained in the core may be released. In oneembodiment the osmogen is a salt such as but not limited to sodiumchloride, potassium chloride, sodium citrate, or potassium citrate.

The carrier may contain one or more suitable excipients for theformulation of tablets. Examples of suitable excipients include, but arenot limited to, fillers, adsorbents, binders, disintegrants, lubricants,glidants, release-modifying excipients, superdisintegrants,antioxidants, and mixtures thereof.

Suitable fillers include, but are not limited to, watersolublecompressible carbohydrates such as sugars (e.g., dextrose, sucrose,maltose, and lactose), starches (e.g., corn starch), sugar-alcohols(e.g., mannitol, sorbitol, maltitol, erythritol, and xylitol), starchhydrolysates (e.g., dextrins, and maltodextrins), and water insolubleplastically deforming materials (e.g., microcrystalline cellulose orother cellulosic derivatives), and mixtures thereof. Suitable adsorbents(e.g., to adsorb the liquid drug composition) include, but are notlimited to, water-insoluble adsorbents such as dicalcium phosphate,tricalcium phosphate, silicified microcrystalline cellulose (e.g., suchas distributed under the PROSOLV brand (PenWest Pharmaceuticals,Patterson, N.Y.)), magnesium aluminometasilicate (e.g., such asdistributed under the NEUSILIN™ brand (Fuji Chemical Industries (USA)Inc., Robbinsville, N.J.), clays, silicas, bentonite, zeolites,magnesium silicates, hydrotalcite, veegum, and mixtures thereof.

Suitable binders include, but are not limited to, dry binders such aspolyvinyl pyrrolidone and hydroxypropylmethylcellulose; wet binders suchas water-soluble polymers, including hydrocolloids such as acacia,alginates, agar, guar gum, locust bean, carrageenan,carboxymethylcellulose, tara, gum arabic, tragacanth, pectin, xanthan,gellan, gelatin, maltodextrin, galactomannan, pusstulan, laminarin,scleroglucan, inulin, whelan, rhamsan, zooglan, methylan, chitin,cyclodextrin, chitosan, polyvinyl pyrrolidone, cellulosics, sucrose, andstarches; and mixtures thereof. Suitable disintegrants include, but arenot limited to, sodium starch glycolate, cross-linkedpolyvinylpyrrolidone, cross-linked carboxymethylcellulose, starches,microcrystalline cellulose, and mixtures thereof.

Suitable lubricants include, but are not limited to, long chain fattyacids and their salts, such as magnesium stearate and stearic acid,talc, glycerides waxes, and mixtures thereof. Suitable glidants include,but are not limited to, colloidal silicon dioxide. Suitablerelease-modifying excipients include, but are not limited to, insolubleedible materials, pH-dependent polymers, and mixtures thereof.

Suitable insoluble edible materials for use as release-modifyingexcipients include, but are not limited to, water-insoluble polymers andlow-melting hydrophobic materials, copolymers thereof, and mixturesthereof. Examples of suitable water-insoluble polymers include, but arenot limited to, ethylcellulose, polyvinyl alcohols, polyvinyl acetate,polycaprolactones, cellulose acetate and its derivatives, acrylates,methacrylates, acrylic acid copolymers, copolymers thereof, and mixturesthereof. Suitable low-melting hydrophobic materials include, but are notlimited to, fats, fatty acid esters, phospholipids, waxes, and mixturesthereof. Examples of suitable fats include, but are not limited to,hydrogenated vegetable oils such as for example cocoa butter,hydrogenated palm kernel oil, hydrogenated cottonseed oil, hydrogenatedsunflower oil, and hydrogenated soybean oil, free fatty acids and theirsalts, and mixtures thereof. Examples of suitable fatty acid estersinclude, but are not limited to, sucrose fatty acid esters, mono-, di-,and triglycerides, glyceryl behenate, glyceryl palmitostearate, glycerylmonostearate, glyceryl tristearate, glyceryl trilaurylate, glycerylmyristate, GlycoWax-932, lauroyl macrogol-32 glycerides, stearoylmacrogol-32 glycerides, and mixtures thereof. Examples of suitablephospholipids include phosphotidyl choline, phosphotidyl serene,phosphotidyl enositol, phosphotidic acid, and mixtures thereof. Examplesof suitable waxes include, but are not limited to, carnauba wax,spermaceti wax, beeswax, candelilla wax, shellac wax, microcrystallinewax, and paraffin wax; fat-containing mixtures such as chocolate, andmixtures thereof. Examples of super disintegrants include, but are notlimited to, croscarmellose sodium, sodium starch glycolate andcross-linked povidone (crospovidone). In one embodiment the tablet corecontains up to about 5 percent by weight of such super disintegrant.

Examples of antioxidants include, but are not limited to, tocopherols,ascorbic acid, sodium pyrosulfite, butylhydroxytoluene, butylatedhydroxyanisole, edetic acid, and edetate salts, and mixtures thereof.Examples of preservatives include, but are not limited to, citric acid,tartaric acid, lactic acid, malic acid, acetic acid, benzoic acid, andsorbic acid, and mixtures thereof.

The osmotic tablets of the present invention include an osmotic coating.An osmotic coating is one that is semipermeable thereby allows water tobe drawn into the tablet core, e.g., for the purposes of releasing theactive ingredient such as through a pre-made hole in the coating orthrough coating itself it is semipermeable membrane. The osmoticcoating, thus, does not fully dissolve upon contact with water In oneembodiment, the osmotic coating contains a water soluble component suchas a water solible film former which aids in facilitating a furtherinflux of water upon contact with water. In the current invention theosmotic coating is applied via spray coating. Suitable spray coatingtechniques include spray coating via a coating pan or fluid bed processsuch as Wurster coating or top spray fluid bed coating as described inthe text, “The Theory and Practice of Industrial Pharmacy”, Lachman,Leon et. al, 3rd ed. The osmotic coating may be applied using a solutionprepared with water, organic solvents, or mixtures thereof. Suitableorganic solvents include but are not limited to acetone, isopropanol,methylene chloride, hexane, methanol, ethanol, and mixtures thereof. Inone embodiment the polymer(s) are dissolved in the coating solution. Inone embodiment, the polymer(s) are dispersed, as is the case whenapplying water insoluble polymers via a dispersion or as is the casewhen using ethylcellulose dispersions.

In one embodiment in which the osmotic coating functions as asemipermeable membrane (e.g., allowing water or solvent to pass into thecore, but being impermeable to dissolved pharmaceutically active agent,thereby preventing the passage of pharmaceutically active agenttherethrough) the film former is selected from water insoluble polymers,pH-dependent polymers, water soluble polymers, and combinations thereof.In one embodiment, the osmotic coating includes a water insolublepolymer and a pore forming material. Examples of suitablewater-insoluble polymers include ethylcellulose, polyvinyl alcohols,polyvinyl acetate, polycaprolactones, cellulose acetate and itsderivatives, acrylates, methacrylates, acrylic acid copolymers, andcombinations thereof. In one embodiment, the water insoluble polymer iscellulose acetate. In one embodiment, the osmotic coating includes fromabout 10 to about 100 weight percent of a water insoluble film former.

In one embodiment of the osmotic coating, the water insoluble polymer iscombined with a water soluble film former in order to create pores inthe resulting semi-permeable membrane. Examples of suitable film formersinclude, but are not limited to: water soluble vinyl polymers such aspolyvinylalcohol (PVA); water soluble polycarbohydrates such ashydroxypropyl starch, hydroxyethyl starch, pullulan, methylethyl starch,carboxymethyl starch, pre-gelatinized starches, and film-formingmodified starches; water swellable cellulose derivatives such ashydroxypropyl cellulose (HPC), hydroxypropylmethyl cellulose (HPMC),methyl cellulose (MC), hydroxyethylmethylcellulose (HEMC),hydroxybutylmethylcellulose (HBMC), hydroxyethylethylcellulose (HEEC),and hydroxyethylhydroxypropylmethyl cellulose (HEMPMC); water solublecopolymers such as methacrylic acid and methacrylate ester copolymers,polyvinyl alcohol and polyethylene glycol copolymers, polyethylene oxideand polyvinylpyrrolidone copolymers; and mixtures thereof.

In one embodiment, a pH dependent polymer is incorporated into theosmotic coating. In one embodiment, the pH dependent polymer is used ata level of from about 10 to about 50 percent by weight of the osmoticcoating. Suitable film-forming pH-dependent polymers include, but arenot limited to, enteric cellulose derivatives, such as for examplehydroxypropyl methylcellulose phthalate, hydroxypropyl methylcelluloseacetate succinate, and cellulose acetate phthalate; natural resins suchas shellac and zein; enteric acetate derivatives such aspolyvinylacetate phthalate, cellulose acetate phthalate, andacetaldehyde dimethylcellulose acetate; and enteric acrylate derivativessuch as for example polymethacrylate-based polymers such aspoly(methacrylic acid, methyl methacrylate) 1:2 (commercially availablefrom Rohm Pharma GmbH under the tradename EUDRAGIT STM), andpoly(methacrylic acid, methyl methacrylate) 1:1 (commercially availablefrom Rohm Pharma GmbH under the tradename EUDRAGIT LTM); andcombinations thereof. In one embodiment, the osmotic coating has anaverage thickness of at least 5 microns, such as from about 10 micronsto about 200 microns, e.g. from about 20 microns to about 150 microns,e.g. from about 30 to about 150 microns. In one embodiment, the osmoticcoating is free of porosity (e.g., wherein the pore volume is in a porediameter range of less than 0.01 g/cc). In one embodiment, the averagepore diameter of the osmotic coating is less than about 0.2 microns(e.g., less than about 0.15 microns).

In one embodiment, the osmotic coating is substantially free ofanpharmaceutically active agent. In one embodiment the osmotic coatingincludes anpharmaceutically active agent which is different than thepharmaceutically active agent included in the immediate release coating.In one embodiment, the osmotic coating includes a plasticizer. In oneembodiment the plasticizer must be of sufficient quantity to withstandthe compression force of the immediate release coating. Suitableplasticizers include, but are not limited to: polyethylene glycol;propylene glycol; glycerin; sorbitol; triethyl citrate; tributylcitrate; dibutyl sebecate; vegetable oils such as castor oil, grape oil,olive oil, and sesame oil; surfactants such as polysorbates, sodiumlauryl sulfates, and dioctyl-sodium sulfosuccinates; mono acetate ofglycerol; diacetate of glycerol; triacetate of glycerol; natural gums;triacetin; acetyltributyl citrate; diethyloxalate; diethylmalate;diethyl fumarate; diethylmalonate; dioctylphthalate; dibutylsuccinate;glycerol tributyrate; hydrogenated castor oil; fatty acids such aslauric acid; glycerides such as mono-, di-, and/or triglycerides, whichmay be substituted with the same or different fatty acids groups suchas, for example, stearic, palmitic, and oleic and the like; and mixturesthereof. In one embodiment, the plasticizer is triethyl citrate.

In one embodiment, at least about 50 percent of the cross-sectional areaof the osmotic coating used in tablets of this invention is striated,such as at least about 80% of the cross-sectional area of the osmoticcoating portion is striated. As used herein, “striated” meansnon-homogeneous with respect to appearance and with respect to theinternal structure of the coating portion when viewed under anymagnification and lighting conditions, at which point striations orlayers can be viewed. Compressed portions of a pharmaceutical oraldosage forms do not display striated areas, wherein spray coatedportions display striations. For example a crosssection of the osmoticcoating portion is striated, and nonuniform with respect to refractiveproperties when observed utilizing a light microscope or a scanningelectron microscope at a magnification of about 50 to about 400 times.The characteristic striations are indicative of the spray-coatingprocess consisting of multiple repetitions of the steps consisting of:(a) application via spraying of coating solution; followed by (b) warmair drying, to a tumbling bed of tablets in a revolving coating pan suchthat numerous layers of coating material are built up as eachapplication of coating material dries to form a layer. In oneembodiment, the thickness of an individual striated layer is the rangeof about 10 microns to about 15 microns.

In certain embodiments, the osmotic coating is semipermeable (e.g.,containing a plurality of small opening) and does not require theaddition of an additional opening via laser or other means. In one suchembodiment, the semi-permeable membrane of the osmotic coating alsoallows for the release of the active ingredient in the tablet corethrough the membrane in a zero-order or first-order release manner.

In one embodiment, the immediate release coating has an averagethickness of at least 50 microns, such as from about 50 microns to about2500 microns; e.g., from about 250 microns to about 1000 microns. Inembodiment, the immediate release coating is typically compressed at adensity of more than about 0.9 g/cc, as measured by the weight andvolume of that specific layer.

In one embodiment, the immediate release coating contains a firstportion and a second portion, wherein at least one of the portionscontains the second pharmaceutically active agent. In one embodiment,the portions contact each other at a center axis of the tablet. In oneembodiment, the first portion includes the first pharmaceutically activeagent and the second portion includes the second pharmaceutically activeagent.

In one embodiment, the first portion contains the first pharmaceuticallyactive agent and the second portion contains the second pharmaceuticallyactive agent. In one embodiment, one of the portions contains a thirdpharmaceutically active agent. In one embodiment one of the portionscontains a second immediate release portion of the same pharmaceuticallyactive agent as that contained in the tablet core.

In one embodiment, the outer coating portion is prepared as a dry blendof materials prior to addition to the coated tablet core. In anotherembodiment the outer coating portion is included of a dried granulationincluding the pharmaceutically active agent.

In one embodiment, a suitable flavor or aroma agent may be added to theouter coating. Examples of suitable flavor and aroma agents include, butare not limited to, essential oils including distillations, solventextractions, or cold expressions of chopped flowers, leaves, peel orpulped whole fruit containing mixtures of alcohols, esters, aldehydesand lactones; essences including either diluted solutions of essentialoils, or mixtures of synthetic chemicals blended to match the naturalflavor of the fruit (e.g., strawberry, raspberry, and black currant);artificial and natural flavors of brews and liquors (e.g., cognac,whisky, rum, gin, sherry, port, and wine); tobacco, coffee, tea, cocoa,and mint; fruit juices including expelled juice from washed, scrubbedfruits such as lemon, orange, and lime; mint; ginger; cinnamon;cacoe/cocoa; vanilla; liquorice; menthol; eucalyptus; aniseeds nuts(e.g., peanuts, coconuts, hazelnuts, chestnuts, walnuts, and colanuts);almonds; raisins; and powder, flour, or vegetable material partsincluding tobacco plant parts (e.g., the genus Nicotiana in amounts notcontributing significantly to a level of therapeutic nicotine), andmixtures thereof.

formulations with different drug release mechanisms described abovecould be combined in a final dosage form containing single or multipleunits. Examples of multiple units include multilayer tablets, capsulescontaining tablets, beads, or granules in a solid or liquid form.Typical, immediate release formulations include compressed tablets,gels, films, coatings, liquids and particles that can be encapsulated,for example, in a gelatin capsule. Many methods for preparing coatings,covering or incorporating drugs, are known in the art.

The immediate release dosage, unit of the dosage form, i.e., a tablet, aplurality of drug-containing beads, granules or particles, or an outerlayer of a coated core dosage form, contains a therapeutically effectivequantity of the active agent with conventional pharmaceuticalexcipients. The immediate release dosage unit may or may not be coated,and may or may not be admixed with the delayed release dosage unit orunits (as in an encapsulated mixture of immediate releasedrug-containing granules, particles or beads and delayed releasedrug-containing granules or beads). A preferred method for preparingimmediate release tablets (e.g., as incorporated into a capsule) is bycompressing a drugcontaining blend, e.g., blend of granules, preparedusing a direct, blend, wet-granulation or dry-granulation process.Immediate release tablets may also be molded rather than compressed,starting with a moist material containing a suitable water-solublelubricant. However, preferred tablets described herein are manufacturedusing compression rather than molding. A preferred method for formingimmediate release drug-containing blend is to mix drug particlesdirectly with one or more excipients such as diluents (or fillers),binders, disintegrants, lubricants, glidants, and/or colorants. As analternative to direct blending, a drug-containing blend may be preparedby using a wet-granulation or dry-granulation process. Beads containingthe active agent may also be prepared by any one of a number ofconventional techniques, typically starting from a fluid dispersion. Forexample, a typical method for preparing drug-containing beads involvesblending the active agent with conventional pharmaceutical excipientssuch as microcrystalline cellulose, starch, polyvinylpyrrolidone,methylcellulose, talc, metallic stearates, and silicone dioxide. Theadmixture is used to coat a bead core such as a sugar sphere (e.g.,“non-parcil”) having a size of approximately 20 to 60 mesh.

An alternative procedure for preparing drug beads is by blending tiledrug with one or more pharmaceutically acceptable excipients, such asmicrocrystalline cellulose, lactose, cellulose, polyvinyl pyrrolidone,talc, magnesium stearate, and a disintegrant, extruding the blend,spheronizing the extrudate, drying and optionally coating the bead toform immediate release beads.

Extended release formulations are generally prepared as diffusion orosmotic systems, for example, as described in “Remington—The Science andPractice of Pharmacy”, 20th. Ed., Lippincott Williams & Wilkins,Baltimore, Md., 2000). A diffusion system typically consists of one oftwo types of devices, reservoir and matrix, which are wellknown anddescribed in die art. The matrix devices are generally prepared bycompressing the drug with a slowly dissolving polymer carrier into atablet form. The three major types of materials used in the preparationof matrix devices are insoluble plastics, hydrophilic polymers, andfatty compounds. Plastic matrices include, but are not limited to,methyl acrylate-methyl methacrylate, polyvinyl chloride, andpolyethylene. Hydrophilic polymers include, but are not limited to,methylcellulose, hydroxypropylcellulose, hydorxypropylmethylcellulose,sodium carboxymethylcellulose, and Carbopol™ 934, and polyethyleneoxides. Fatty compounds include, but are not limited to, various waxessuch as carnauba wax and glyceryl tristearate. Alternatively, extendedrelease formulations can be prepared using osmotic systems or byapplying a semi-permeable coating to the dosage form. In the lattercase, the desired drug release profile can be achieved by combining, lowpermeability and high permeability coating materials in suitableproportion.

An immediate release portion can be added to the extended release systemby means of either applying an immediate release layer on top of theextended release core; using coating or compression processes or in amultiple unit system such as a capsule containing extended and immediaterelease beads.

Extended release tablets containing hydrophilic polymers are prepared bytechniques commonly known in the art such as direct compression, wetgranulation, or dry granulation processes. These formulations usuallyincorporate polymers, diluents, binders, and lubricants as well as theactive pharmaceutical ingredient. The usual diluents include inertpowdered substances such as different kinds of starch, powdered,cellulose, especially crystalline and microcrystalline cellulose, sugarssuch as fructose, mannitol and sucrose, grain flours and similar ediblepowders. Typical diluents include, for example, various types of starch,lactose, mannitol, kaolin, calcium phosphate or sulfate, inorganic saltssuch as sodium chloride and powdered sugar. Powdered cellulosederivatives are also useful. Typical tablet binders include substancessuch as starch, gelatin and sugars such as lactose, fructose, andglucose. Natural and synthetic gums, including acacia, alginates,methylcellulose, and polyvinylpyrrolidine can also be used. Polyethyleneglycol, hydrophilic polymers, ethycellulose and waxes can also serve asbinders. A lubricant is necessary in a tablet formulation to prevent thetablet and punches from sticking in the die. The lubricant is chosenfrom such slippery solids as tale, magnesium and calcium stearate,stearic acid and hydrogenated vegetable oils. Extended release tabletscontaining wax materials are generally prepared using methods known inthe art such as a direct blend method, a congealing method, and anaqueous dispersion method. In the congealing method, the drug is mixedwith a wax material and either spray-congealed or congealed and screenedand processed.

Delayed release dosage formulations are created by coating a soliddosage form with a film of a polymer which is insoluble in the acidenvironment of the stomach, but soluble in the neutral environment ofsmall intestines. The delayed release dosage units can be prepared, forexample, by coating a drug or a drug-containing composition with aselected coating material. The drug-containing composition may be atablet for incorporation into a capsule, a tablet for use as an innercore in a “coated core” dosage form, or a plurality of drug-containingbeads, particles or granules, for incorporation into either a tablet orcapsule. Preferred coating materials include bioerodible, graduallyhydrolyzable, gradually water-soluble, and/or enzymatically degradablepolymers, and may be conventional “enteric” polymers. Enteric polymers,as will be appreciated by those skilled in the art, become soluble inthe higher pH environment of the lower gastrointestinal tract or slowlyerode as the dosage form passes through the gastrointestinal tract,while enzymatically degradable polymers are degraded by bacterialenzymes present in the lower gastrointestinal tract, particularly in thecolon. Suitable coating materials for effecting delayed release include,but are not limited to, cellulosic polymers such as hydroxypropylcellulose, hydoxyethyl cellulose, hydroxymethyl cellulose, hydroxypropylmethyl cellulose, hydroxypropyl methyl cellulose acetate succinate,hydroxypropylmethyl cellulose phthalate, methylcellulose, ethylcellulose, cellulose acetate, cellulose acetate phthalate, celluloseacetate trimellitate and carboxymethylcellulose sodium; acrylic acidpolymers and copolymers, preferably formed from acrylic acid,methacrylic acid, methyl acrylate, ethyl acrylate, methyl methacrylateand/or ethyl methacrylate, and other methacrylic resins that arecommercially available under the tradename EUDRAGIT™ (Rohm Pharma;[0086] Westerstadt, Germany), including EUDRAGIT™ L30D-55 and L1OO-55(soluble at pH 5,5 and above). EUDRAGIT™ 1,100D (soluble at pH 6.0 andabove), EUDRAGIT™ S (soluble at pH 7.0 and above, as a result of ahigher degree of esterification), and EUDRAGIT™ NE, RL and RS(water-insoluble polymers having different degrees of permeability andexpandability); vinyl polymers and copolymets such as polyvinylpyrrolidone, vinyl acetate, vinylacetate phthalate, vinylacetatecrotonic acid copolymer, and ethylene-vinyl acetate copolymer;enzymatically degradable polymers such as azo polymers, pectin,chitosan, amylase and guar gum; zein and shellac. Combinations ofdifferent coating, materials may also be used. Multi-layer coatingsusing different polymers may also be applied. The preferred coatingweights for particular coating materials may be readily determined bythose skilled in the art by evaluating individual release profiles fortablets, beads and granules prepared with different quantities ofvarious coating materials. It is the combination of materials, method,and form of application that produce the desired releasecharacteristics, which one can determine only from the clinical studies.

The coating composition may include conventional additives, such asplasticizers, pigments, colorants, stabilizing agents, glidants, etc. Aplasticizer is normally present to reduce the fragility of the coating,and will generally represent about 10 wt. % to 50 wt. % relative to thedry weight of the polymer. Examples of typical plasticizers includepolyethylene glycol, propylene glycol, triacetin, dimethyl phthalate,diethyl phthalate, dibutyl phthalate, dibutyl sebacate, triethylcitrate, tributyl citrate, triethyl acetyl citrate, castor oil andacetylated monoglycerides. A stabilizing agent is preferably used tostabilize particles in the dispersion. Typical stabilizing agents arenonionic emulsifiers such as sorbitan esters, polysorbates andpolyvinylpyrrolidone. Glidants are recommended to reduce stickingeffects during film formation and drying, and will generally representapproximately 25 wt. % to 100 wt. % of the polymer weight in the coatingsolution. One effective glidant is talc. Other glidants such asmagnesium stearate and glycerol monostearates may also be used. Pigmentssuch as titanium dioxide may also be used. Small quantities of ananti-foaming agent, such as a silicone (e.g., simethicone), may also beadded to the coating composition.

Alternatively, a delayed release tablet may be formulated by dispersingtire drug within a matrix of a suitable material such as a hydrophilicpolymer or a fatty compound. Suitable hydrophilic polymers include, butare not limited to, polymers or copolymers of cellulose, celluloseester, acrylic acid, methacrylic acid, methyl acrylate, ethyl acrylate,and vinyl or enzymatically degradable polymers or copolymers asdescribed above. These hydrophilic polymers are particularly useful forproviding a delayed release matrix. Fatty compounds for use as a matrixmaterial include, but are hot limited to, waxes (e.g. carnauba wax) andglycerol tristearate. Once the active ingredient is mixed with thematrix material, the mixture can be compressed into tablets.

A pulsed release dosage form is one that mimics a multiple dosingprofile without repeated dosing and typically allows at least a twofoldreduction in dosing frequency as compared to the drug presented as aconventional dosage form (e.g., as a solution or prompt drug-releasing,conventional solid dosage form). A pulsed release profile ischaracterized by a time period of no release (lag time) or reducedrelease followed by rapid drug release.

Each dosage form contains a therapeutically effective amount of activeagent. In one embodiment of dosage forms that mimic a twice daily dosingprofile, approximately 30 wt. % to 70 wt. %, preferably 40 wt. % to 60wt. %, of the total amount of active agent in the dosage form isreleased in the initial pulse, and, correspondingly approximately 70 wt.% to 3.0 wt. %, preferably 60 wt. % to 40 wt. %, of the total amount ofactive agent in the dosage form is released in the second pulse. Fordosage forms mimicking the twice daily dosing profile, the second pulseis preferably released approximately 3 hours to less than 14 hours, andmore preferably approximately 5 hours to 12 hours, followingadministration.

For dosage forms mimicking a three times daily dosing profile,approximately 25 wt. % to 40 wt. % of the total amount of active agentin the dosage form is released in the initial pulse, and approximately25 wt. % to 40 wt. % of the total amount of active agent in the dosageform is released in each of the second and third pulses. For dosageforms that mimic a three times daily dosing profile, release of thesecond pulse preferably takes place approximately 3 hours to 10 hours,and more preferably approximately 4 to 9 hours, following oraladministration. Release of the third pulse occurs about 2 hours to about8 hours following the second pulse, which is typically about 5 hours toapproximately 18 hours following oral administration.

The dosage form can be a closed capsule housing at least twodrug-containing dosage units, each dosage unit containing one or morecompressed tablets, or may contain, a plurality of beads, granules orparticles, providing that each dosage unit has a different drug releaseprofile. The immediate release dosage unit releases drug substantiallyimmediately following oral administration to provide an initial dose.The delayed release dosage unit releases drug approximately 3 hours to14 hours following oral administration to provide a second dose.Finally, an optional second delayed release dosage unit releases drugabout 2 hours to 8 hours following the release of the second dose, whichis typically 5 hours to 18 hours following oral administration.

Another dosage form contains a compressed tablet or a capsule having adrug-containing immediate release dosage unit, a delayed release dosageunit and an optional second delayed release dosage unit. In this dosageform, the immediate release dosage unit contains a plurality of beads,granules particles that release drug substantially immediately followingoral administration to provide an initial dose. The delayed releasedosage unit contains a plurality of coated beads or granules, whichrelease drug approximately 3 hours to 14 hours following oraladministration to provide a second dose.

An optional second delayed release dosage unit contains coated beads orgranules that release drug about 2 to 8 hours following administrationof the initial delayed release dose, which is typically 5 to 18 hoursfollowing oral administration. The beads or granules in the delayedrelease dosage unites) are coated with a bioerodible polymeric material.This coating prevents the drug from being released until the appropriatetime, i.e., approximately 3 hours to less than 14 hours following oraladministration for the delayed release dosage unit and at least 5 hoursto approximately 18 hours following oral administration for the optionalsecond delayed release dosage unit. In this dosage form the componentsmay be admixed in the tablet or may be layered to form a laminatedtablet.

Another dosage form is a tablet having a drug-containing immediaterelease dosage unit, a delayed release dosage unit, and an optionalsecond delayed release dosage unit, wherein the immediate release dosageunit comprises an outer layer that releases the drug substantiallyimmediately following oral administration. The arrangement of theremaining delayed release dosage(s), however, depends upon whether thedosage form is designed to mimic twice daily dosing or three times dailydosing.

In the dosage form mimicking twice daily dosing, the delayed releasedosage unit contains an inner core that is coated with a bioerodiblepolymeric material. The coating is applied such that release of the drugoccurs approximately 3 hours to less than 14 hours following oraladministration. In this form, the outer layer completely surrounds theinner core. In the dosage form mimicking three times a day dosing, the(first) delayed release dose contains an internal layer that releasesdrug approximately 3 hours to less than 14 hours following oraladministration. This internal layer is surrounded by the outer layer.The second delayed release dosage unit generally contains an inner corethat releases the drug at least 5 hours to approximately 18 hoursfollowing oral administration. Thus, the layers of this tablet (startingfrom the external surface) contain an outer layer, an internal layer andan inner core. The inner core contains delayed release beads orgranules. Furthermore, the internal layer contains the drug coated witha bioerodible polymeric material. Alternatively, in this particulardosage form mimicking three times a day dosing, both the delayed releasedosage unit and second delayed release dosage units are surrounded by aninner layer. This inner layer is free of active agent. Thus, the layersof this tablet (starting from the external surface) comprise an outerlayer, inner layer and an admixture of the delayed release dosage units.The first delayed release pulse occurs once the inner layer issubstantially eroded thereby releasing the admixture of the delayedrelease dosage units. The dose corresponding to the (first) delayedrelease dosage unit is released immediately since the inner layer hasprevented access to this dose for the appropriate time, e.g., fromapproximately 3 hours to 10 hours. The second delayed release dose,however, is formulated to effectively delay release for at least 5 hoursto approximately 18 hours following oral administration.

For formulations mimicking twice daily dosing, it is preferred that thedelayed release dose is released approximately 3 hours to up to 14hours, more preferably approximately 5 hours to up to 12 hours,following oral administration. For formulations mimicking three timesdaily dosing, it is preferred that the (first) delayed release dose isreleased approximately 3 to 10 hours, preferably 4 hours to 9 hours,following oral administration. For dosage forms containing a third dose,the third dose (i.e., the second delayed release dose) is released atleast 5 hours to approximately 18 hours following oral administration.

In still another embodiment, a dosage form is provided which contains acoated core-type delivery system wherein the outer layer contains animmediate release dosage unit containing an active agent, such that theactive agent therein is immediately released following oraladministration; an intermediate layer there under which surrounds acore; and a core which contains immediate release beads or granules anddelayed release beads or granules, such that the second dose is providedby the immediate release beads or granules and the third dose isprovided by the delayed release beads or granules.

For purposes of transdermal (e.g., topical) administration, dilutesterile, aqueous or partially aqueous solutions (usually in about 0.1%to 5% concentration), otherwise similar to the above parenteralsolutions, may be prepared.

Methods of preparing various pharmaceutical compositions with a certainamount of one or more compounds of Formula I, Formula II, Formula III,Formula IV, Formula V or Formula VI or other active agents are known, orwill be apparent in light of this disclosure, to those skilled in thisart. For examples of methods of preparing pharmaceutical compositions,see Remington's Pharmaceutical Sciences, Mack Publishing Company,Easton, Pa., 19th Edition (1995).

In addition, in certain embodiments, subject compositions of the presentapplication maybe lyophilized or subjected to another appropriate dryingtechnique such as spray drying. The subject compositions may beadministered once, or may be divided into a number of smaller doses tobe administered at varying intervals of time, depending in part on therelease rate of the compositions and the desired dosage.

formulations useful in the methods provided herein include thosesuitable for oral, nasal, topical (including buccal and sublingual),rectal, vaginal, aerosol and/or parenteral administration. Theformulations may conveniently be presented in unit dosage form and maybe prepared by any methods well known in the art of pharmacy. The amountof a subject composition which may be combined with a carrier materialto produce a single dose may vary depending upon the subject beingtreated, and the particular mode of administration.

Methods of preparing these formulations or compositions include the stepof bringing into association subject compositions with the carrier and,optionally, one or more accessory ingredients. In general, theformulations are prepared by uniformly and intimately bringing intoassociation a subject composition with liquid carriers, or finelydivided solid carriers, or both, and then, if necessary, shaping theproduct.

The compounds of Formula I, Formula II, Formula III, Formula IV, FormulaV or Formula VI described herein may be administered in inhalant oraerosol formulations. The inhalant or aerosol formulations may compriseone or more agents, such as adjuvants, diagnostic agents, imagingagents, or therapeutic agents useful in inhalation therapy. The finalaerosol formulation may for example contain 0.005-90% w/w, for instance0.005-50%, 0.005-5% w/w, or 0.01-1.0% w/w, of medicament relative to thetotal weight of the formulation.

It is desirable, but by no means required, that the formulations hereincontain no components which may provoke the degradation of stratosphericozone. In particular it is desirable that the formulations aresubstantially free of chlorofluorocarbons such as CC13F, CC12F2 andCF3CC13. As used to refer to ozone-damaging agents, “substantially free”means less than 1% w/w based upon the propellant system, in particularless than 0.5%, for example 0.1% or less.

The propellant may optionally contain an adjuvant having a higherpolarity and/or a higher boiling point than the propellant. Polaradjuvants which may be used include (e.g., C2-6) aliphatic alcohols andpolyols such as ethanol, isopropanol and propylene glycol. In general,only small quantities of polar adjuvants (e.g., 0.05-3.0% w/w) may berequired to improve the stability of the dispersion—the use ofquantities in excess of 5% w/w may tend to dissolve the medicament. Theformulations described herein may contain less than 1% w/w, e.g., about0.1% w/w, of polar adjuvant. However, the formulations may besubstantially free of polar adjuvants, such as ethanol. Suitablevolatile adjuvants include saturated hydrocarbons such as propane,n-butane, isobutane, pentane and isopentane and alkyl ethers such asdimethyl ether. In general, up to 50% w/w of the propellant may comprisea volatile adjuvant, for example 1 to 30% w/w of a volatile saturatedC1-C6 hydrocarbon.

Optionally, the aerosol formulations may further comprise one or moresurfactants. The surfactants must be physiologically acceptable uponadministration by inhalation. Within this category are includedsurfactants such as L-α-phosphatidylcholine (PC),1,2-dipalmitoylphosphatidycholine (DPPC), oleic acid, sorbitantrioleate, sorbitan mono-oleate, sorbitan monolaurate, polyoxyethylene(20) sorbitan monolaurate, polyoxyethylene (20) sorbitan monooleate,natural lecithin, oleyl polyoxyethylene (2) ether, stearylpolyoxyethylene (2) ether, lauryl polyoxyethylene (4) ether, blockcopolymers of oxyethylene and oxypropylene, synthetic lecithin,diethylene glycol dioleate, tetrahydrofurfuryl oleate, ethyl oleate,isopropyl myristate, glyceryl monooleate, glyceryl monostearate,glyceryl monoricinoleate, cetyl alcohol, stearyl alcohol, polyethyleneglycol 400, cetyl pyridinium chloride, benzalkonium chloride, olive oil,glyceryl monolaurate, corn oil, cotton seed oil, and sunflower seed oil.Appropriate surfactants include lecithin, oleic acid, and sorbitantrioleate.

Ophthalmic formulations, eye ointments, powders, solutions and the like,are also contemplated as being within the scope of the disclosuresherein.

Certain pharmaceutical compositions disclosed herein suitable forparenteral administration comprise one or more subject compositions incombination with one or more pharmaceutically acceptable sterile,isotonic, aqueous, or non-aqueous solutions, dispersions, suspensions oremulsions, or sterile powders which may be reconstituted into sterileinjectable solutions or dispersions just prior to use, which may containantioxidants, buffers, bacteriostats, solutes which render theformulation isotonic with the blood of the intended recipient orsuspending or thickening agents.

Examples of suitable aqueous and non-aqueous carriers which may beemployed in the pharmaceutical compositions include water, ethanol,polyols (such as glycerol, propylene glycol, polyethylene glycol, andthe like), and suitable mixtures thereof, vegetable oils, such as oliveoil, and injectable organic esters, such as ethyl oleate. Properfluidity may be maintained, for example, by the use of coatingmaterials, such as lecithin, by the maintenance of the required particlesize in the case of dispersions, and by the use of surfactants.

formulations suitable for oral administration may be in the form ofcapsules, cachets, pills, tablets, lozenges (using a flavored basis,usually sucrose and acacia or tragacanth), powders, granules, or as asolution or a suspension in an aqueous or non-aqueous liquid, or as anoil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup,or as pastilles (using an inert base, such as gelatin and glycerin, orsucrose and acacia), each containing a predetermined amount of a subjectcomposition as an active ingredient. Subject compositions may also beadministered as a bolus, electuary, or paste.

In solid dosage forms for oral administration (capsules, tablets, pills,dragees, powders, granules and the like), the subject composition ismixed with one or more pharmaceutically acceptable carriers and/or anyof the following: (1) fillers or extenders, such as starches, lactose,sucrose, glucose, mannitol, and/or silicic acid; (2) binders, such as,for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and/or acacia; (3) humectants, such as glycerol;(4) disintegrating agents, such as agar-agar, calcium carbonate, potatoor tapioca starch, alginic acid, certain silicates, and sodiumcarbonate; (5) solution retarding agents, such as paraffin; (6)absorption accelerators, such as quaternary ammonium compounds; (7)wetting agents, such as, for example, acetyl alcohol and glycerolmonostearate; (8) absorbents, such as kaolin and bentonite clay; (9)lubricants, such a talc, calcium stearate, magnesium stearate, solidpolyethylene glycols, sodium lauryl sulfate, and mixtures thereof; and(10) coloring agents. In the case of capsules, tablets and pills, thepharmaceutical compositions may also comprise buffering agents. Solidcompositions of a similar type may also be employed as fillers in softand hard-filled gelatin capsules using lactose or milk sugars, as wellas high molecular weight polyethylene glycols and the like.

A tablet may be made by compression or molding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared using abinder (for example, gelatin or hydroxypropylmethyl cellulose),lubricant, inert diluent, preservative, disintegrant (for example,sodium starch glycolate or cross-linked sodium carboxymethyl cellulose),surface-altering or dispersing agent. Molded tablets may be made bymolding in a suitable machine a mixture of the subject compositionmoistened with an inert liquid diluent. Tablets, and other solid dosageforms, such as dragees, capsules, pills and granules, may optionally bescored or prepared with coatings and shells, such as enteric coatingsand other coatings well known in the pharmaceutical-formulating art.

There has been widespread use of tablets since the latter part of the19th century and the majority of pharmaceutical dosage forms aremarketed as tablets. Major reasons of tablet popularity as a dosage formare simplicity, low cost and the speed of production. Other reasonsinclude stability of drug product, convenience in packaging, shippingand dispensing. To the patient or consumer, tablets offer convenience ofadministration, ease of accurate dosage, compactness, portability,blandness of taste, ease of administration and elegant distinctiveappearance.

Tablets may be plain, film or sugar coated, bisected, embossed, layeredor sustained-release. They can be made in a variety of sizes, shapes andcolors. Tablets may be swallowed, chewed or dissolved in the buccalcavity or beneath the tongue. They may be dissolved in water for localor topical application. Sterile tablets are normally used for parenteralsolutions and for implantation beneath the skin.

In addition to the active or therapeutic ingredients, tablets maycontain a number of inert materials known as excipients. They may beclassified according to the role they play in the final tablet. Theprimary composition may include one or more of a filler, binder,lubricant and glidant. Other excipients which give physicalcharacteristics to the finished tablet are coloring agents, and flavors(especially in the case of chewable tablets). Without excipients mostdrugs and pharmaceutical ingredients cannot be directly-compressed intotablets. This is primarily due to the poor flow and cohesive propertiesof most drugs. Typically, excipients are added to a formulation toimpart good flow and compression characteristics to the material beingcompressed. Such properties are imparted through pretreatment steps,such as wet granulation, slugging, spray drying spheronization orcrystallization.

Lubricants are typically added to prevent the tableting materials fromsticking to punches, minimize friction during tablet compression, andallow for removal of the compressed tablet from the die. Such lubricantsare commonly included in the final tablet mix in amounts usually ofabout 1% by weight.

Other desirable characteristics of excipients include the following:high-compressibility to allow strong tablets to be made at lowcompression forces; impart cohesive qualities to the powdered material;acceptable rate of disintegration; good flow properties that can improvethe flow of other excipients in the Formula; and cohesiveness (toprevent tablet from crumbling during processing, shipping and handling).

There are at least three commercially important processes for makingcompressed tablets: wet granulation, direct compression and drygranulation (slugging or roller compaction). The method of preparationand type of excipients are selected to give the tablet formulation thedesired physical characteristics that allow for the rapid compression ofthe tablets. After compression, the tablets must have a number ofadditional attributes, such as appearance, hardness, disintegratingability and an acceptable dissolution profile. Choice of fillers andother excipients will depend on the chemical and physical properties ofthe drug, behavior of the mixture during processing and the propertiesof the final tablets. Preformulation studies are done to determine thechemical and physical compatibility of the active component withproposed excipients.

The properties of the drug, its dosage forms and the economics of theoperation will determine selection of the best process for tableting.Generally, both wet granulation and direct compression are used indeveloping a tablet.

One formulation comprises the following: a compound of Formula I,Formula II, Formula III, Formula IV, Formula V or Formula VI, and abinder. Examples of pharmaceutically acceptable binders include, but arenot limited to, starches; celluloses and derivatives thereof, e.g.,microcrystalline cellulose, hydroxypropyl cellulose hydroxylethylcellulose and hydroxylpropylmethyl cellulose; sucrose; dextrose; cornsyrup; polysaccharides; and gelatin. The binder, e.g., may be present inan amount from about 1% to about 40% by weight of the composition suchas 1% to 30% or 1% to 25% or 1% to 20%.

Optionally, one, two, three or more diluents can be added to theformulations disclosed herein. Examples of pharmaceutically acceptablefillers and pharmaceutically acceptable diluents include, but are notlimited to, confectioner's sugar, compressible sugar, dextrates,dextrin, dextrose, lactose, mannitol, microcrystalline cellulose,powdered cellulose, sorbitol, sucrose and talc. The filler and/ordiluent, e.g., may be present in an amount from about 15% to about 40%by weight of the composition. In certain embodiments, diluents aremicrocrystalline cellulose which is manufactured by the controlledhydrolysis of alpha-cellulose, obtained as a pulp from fibrous plantmaterials, with dilute mineral acid solutions. Following hydrolysis, thehydrocellulose is purified by filtration and the aqueous slurry is spraydried to form dry, porous particles of a broad size distribution.Suitable microcrystalline cellulose will have an average particle sizeof from about 20 nm to about 200 nm. Microcrystalline cellulose isavailable from several suppliers. Suitable microcrystalline celluloseincludes Avicel PH 101, Avicel PH 102, Avicel PH 103, Avicel PH 105 andAvicel PH 200, manufactured by FMC Corporation. The microcrystallinecellulose may be present in a tablet formulation in an amount of fromabout 25% to about 70% by weight. Another appropriate range of thismaterial is from about 30% to about 35% by weight; yet anotherappropriate range of from about 30% to about 32% by weight. Anotherdiluent is lactose. The lactose may be ground to have an averageparticle size of between about 50 μm and about 500 μm prior toformulating. The lactose may be present in the tablet formulation in anamount of from about 5% to about 40% by weight, and can be from about18% to about 35% by weight, for example, can be from about 20% to about25% by weight.

Optionally one, two, three or more disintegrants can be added to theformulations described herein. Examples of pharmaceutically acceptabledisintegrants include, but are not limited to, starches; clays;celluloses; alginates; gums; cross-linked polymers, e.g., cross-linkedpolyvinyl pyrrolidone, cross-linked calcium carboxymethylcellulose andcross-linked sodium carboxymethylcellulose; soy polysaccharides; andguar gum. The disintegrant, e.g., may be present in an amount from about2% to about 20%, e.g., from about 5% to about 10%, e.g., about 7% aboutby weight of the composition. A disintegrant is also an optional butuseful component of the tablet formulation. Disintegrants are includedto ensure that the tablet has an acceptable rate of disintegration.Typical disintegrants include starch derivatives and salts ofcarboxymethylcellulose. Sodium starch glycolate is one appropriatedisintegrant for this formulation. In certain embodiments, thedisintegrant is present in the tablet formulation in an amount of fromabout 0% to about 10% by weight, and can be from about 1% to about 4% byweight, for instance from about 1.5% to about 2.5% by weight.

Optionally one, two, three or more lubricants can be added to theformulations disclosed herein. Examples of pharmaceutically acceptablelubricants and pharmaceutically acceptable glidants include, but are notlimited to, colloidal silica, magnesium trisilicate, starches, talc,tribasic calcium phosphate, magnesium stearate, aluminum stearate,calcium stearate, magnesium carbonate, magnesium oxide, polyethyleneglycol, powdered cellulose and microcrystalline cellulose. Thelubricant, e.g., may be present in an amount from about 0.1% to about 5%by weight of the composition; whereas, the glidant, e.g., may be presentin an amount from about 0.1% to about 10% by weight. Lubricants aretypically added to prevent the tableting materials from sticking topunches, minimize friction during tablet compression and allow forremoval of the compressed tablet from the die. Such lubricants arecommonly included in the final tablet mix in amounts usually less than1% by weight. The lubricant component may be hydrophobic or hydrophilic.Examples of such lubricants include stearic acid, talc and magnesiumstearate. Magnesium stearate reduces the friction between the die walland tablet mix during the compression and ejection of the tablets. Ithelps prevent adhesion of tablets to the punches and dies. Magnesiumstearate also aids in the flow of the powder in the hopper and into thedie. It has a particle size range of 450-550 microns and a density rangeof 1.00-1.80 g/mL It is stable and does not polymerize within thetableting mix. One lubricant, magnesium stearate may also be employed inthe formulation. In some aspects, the lubricant is present in the tabletformulation in an amount of from about 0.25% to about 6%; alsoappropriate is a level of about 0.5% to about 4% by weight; and fromabout 0.1% to about 2% by weight. Other possible lubricants includetalc, polyethylene glycol, silica and hardened vegetable oils. In anoptional embodiment, the lubricant is not present in the formulation,but is sprayed onto the dies or the punches rather than being addeddirectly to the formulation.

Examples of useful excipients which can optionally be added to thecomposition are described in the Handbook of Pharmaceutical Excipients,3rd edition, Edited by A. H. Kibbe, Published by: AmericanPharmaceutical Association, Washington DC, ISBN: 0-917330-96-X, orHandbook of Pharmaceutical Excipients (4th edition), Edited by Raymond CRowe—Publisher: Science and Practice.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the subject compositions, the liquid dosageforms may contain inert diluents commonly used in the art, such as, forexample, water or other solvents, solubilizing agents and emulsifiers,such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, oils (in particular, cottonseed, corn, peanut, sunflower,soybean, olive, castor, and sesame oils), glycerol, tetrahydrofurylalcohol, polyethylene glycols and fatty acid esters of sorbitan, andmixtures thereof.

Suspensions, in addition to the subject compositions, may containsuspending agents such as, for example, ethoxylated isostearyl alcohols,polyoxyethylene sorbitol, and sorbitan esters, microcrystallinecellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth,and mixtures thereof.

formulations for rectal or vaginal administration may be presented as asuppository, which may be prepared by mixing a subject composition withone or more suitable non-irritating carriers comprising, for example,cocoa butter, polyethylene glycol, a suppository wax, or a salicylate,and which is solid at room temperature, but liquid at body temperatureand, therefore, will melt in the appropriate body cavity and release theencapsulated compound(s) and composition(s). formulations which aresuitable for vaginal administration also include pessaries, tampons,creams, gels, pastes, foams, or spray formulations containing suchcarriers as are known in the art to be appropriate.

Dosage forms for transdermal administration include powders, sprays,ointments, pastes, creams, lotions, gels, solutions, patches, andinhalants. A subject composition may be mixed under sterile conditionswith a pharmaceutically acceptable carrier, and with any preservatives,buffers, or propellants that may be required. For transdermaladministration, the complexes may include lipophilic and hydrophilicgroups to achieve the desired water solubility and transport properties.

The ointments, pastes, creams and gels may contain, in addition tosubject compositions, other carriers, such as animal and vegetable fats,oils, waxes, paraffins, starch, tragacanth, cellulose derivatives,polyethylene glycols, silicones, bentonites, silicic acid, talc and zincoxide, or mixtures thereof. Powders and sprays may contain, in additionto a subject composition, excipients such as lactose, talc, silicicacid, aluminum hydroxide, calcium silicates and polyamide powder, ormixtures of such substances. Sprays may additionally contain customarypropellants, such as chlorofluorohydrocarbons and volatile unsubstitutedhydrocarbons, such as butane and propane.

Methods of delivering a composition or compositions via a transdermalpatch are known in the art. Exemplary patches and methods of patchdelivery are described in U.S. Pat. Nos. 6,974,588, 6,564,093,6,312,716, 6,440,454, 6,267,983, 6,239,180, and 6,103,275.

In one embodiment, a transdermal patch may comprise an outer backingfoil, a matrix and a protective liner wherein a) the composition orcompositions are present in the matrix in a solution (which may beoversaturated), b) the matrix may contain 1 to 5% activated SiO2, and c)the matrix may have a moisture content of less than 0.7%. Moisture-freematrix patches which contain activated silicon dioxide in the matrixshow an enhanced drug release into the skin.

In another embodiment, a transdermal patch may comprise: a substratesheet comprising a composite film formed of a resin compositioncomprising 100 parts by weight of a polyvinyl chloride-polyurethanecomposite and 2-10 parts by weight of astyrene-ethylene-butylene-styrene copolymer, a first adhesive layer onthe one side of the composite film, and a polyalkylene terephthalatefilm adhered to the one side of the composite film by means of the firstadhesive layer, a primer layer which comprises a saturated polyesterresin and is formed on the surface of the polyalkylene terephthalatefilm; and a second adhesive layer comprising a styrene-diene-styreneblock copolymer containing a pharmaceutical agent layered on the primerlayer. A method for the manufacture of the above-mentioned substratesheet comprises preparing the above resin composition molding the resincomposition into a composite film by a calendar process, and thenadhering a polyalkylene terephthalate film on one side of the compositefilm by means of an adhesive layer thereby forming the substrate sheet,and forming a primer layer comprising a saturated polyester resin on theouter surface of the polyalkylene terephthalate film.

The pharmaceutical compositions herein can be packaged to produce a“reservoir type” transdermal patch with or without a rate-limiting patchmembrane. The size of the patch and or the rate limiting membrane can bechosen to deliver the transdermal flux rates desired. Such a transdermalpatch can consist of a polypropylene/polyester impervious backing memberheat-sealed to a polypropylene porous/permeable membrane with areservoir there between. The patch can include a pharmaceuticallyacceptable adhesive (such as a acrylate, silicone or rubber adhesive) onthe membrane layer to adhere the patch to the skin of the host, e.g., amammal such as a human. A release liner such as a polyester releaseliner can also be provided to cover the adhesive layer prior toapplication of the patch to the skin as is conventional in the art. Thispatch assembly can be packaged in an aluminum foil or other suitablepouch, again as is conventional in the art.

Alternatively, the compositions herein can be formulated into a“matrix-type” transdermal patch. Drug Delivery Systems Characteristicsand Biomedical Application, R. L Juliano, ed., Oxford University Press.N.Y. (1980); and Controlled Drug Delivery, Vol. I Basic Concepts,Stephen D. Bruck (1983) describe the theory and application of methodsuseful for transdermal delivery systems. The drug-matrix could be formedutilizing various polymers, e.g. silicone, polyvinyl alcohol. The “drugmatrix” may then be packaged into an appropriate transdermal patch.

Another type of patch comprises incorporating the drug directly in apharmaceutically acceptable adhesive and laminating the drug-containingadhesive onto a suitable backing member, e.g. a polyester backingmembrane. The drug should be present at a concentration which will notaffect the adhesive properties, and at the same time deliver therequired clinical dose.

Transdermal patches may be passive or active. Passive transdermal drugdelivery systems currently available, such as the nicotine, estrogen andnitroglycerine patches, deliver small-molecule drugs. Many of the newlydeveloped proteins and peptide drugs are too large to be deliveredthrough passive transdermal patches and may be delivered usingtechnology such as electrical assist (iontophoresis) for large-moleculedrugs.

Iontophoresis is a technique employed for enhancing the flux of ionizedsubstances through membranes by application of electric current. Oneexample of an iontophoretic membrane is given in U.S. Pat. No. 5,080,646to Theeuwes. The principal mechanisms by which iontophoresis enhancesmolecular transport across the skin are (a) repelling a charged ion froman electrode of the same charge, (b) electroosmosis, the convectivemovement of solvent that occurs through a charged pore in response thepreferential passage of counter-ions when an electric field is appliedor (c) increase skin permeability due to application of electricalcurrent.

In some cases, it may be desirable to administer in the form of a kit,it may comprise a container for containing the separate compositionssuch as a divided bottle or a divided foil packet. Typically the kitcomprises directions for the administration of the separate components.The kit form is particularly advantageous when the separate componentsare preferably administered in different dosage forms (e.g., oral andparenteral), are administered at different dosage intervals, or whentitration of the individual components of the combination is desired bythe prescribing physician.

An example of such a kit is a so-called blister pack. Blister packs arewell known in the packaging industry and are widely used for thepackaging of pharmaceutical unit dosage forms (tablets, capsules, andthe like). Blister packs generally consist of a sheet of relativelystiff material covered with a foil of a plastic material that may betransparent. During the packaging process recesses are formed in theplastic foil. The recesses have the size and shape of the tablets orcapsules to be packed. Next, the tablets or capsules are placed in therecesses and the sheet of relatively stiff material is sealed againstthe plastic foil at the face of the foil which is opposite from thedirection in which the recesses were formed. As a result, the tablets orcapsules are sealed in the recesses between the plastic foil and thesheet. In some embodiments the strength of the sheet is such that thetablets or capsules can be removed from the blister pack by manuallyapplying pressure on the recesses whereby an opening is formed in thesheet at the place of the recess. The tablet or capsule can then beremoved via said opening.

The term “sample” refers to a sample of a body fluid, to a sample ofseparated cells or to a sample from a tissue or an organ. Samples ofbody fluids can be obtained by well known techniques and include,preferably, samples of blood, plasma, serum, or urine, more preferably,samples of blood, plasma or serum. Tissue or organ samples may beobtained from any tissue or organ by, e.g., biopsy. Separated cells maybe obtained from the body fluids or the tissues or organs by separatingtechniques such as centrifugation or cell sorting. Preferably, cell-,tissue- or organ samples are obtained from those cells, tissues ororgans which express or produce the peptides referred to herein.

Methods of Making

The present invention is exemplified by the following non-limitingexamples. Examples of synthetic pathways useful for making compounds ofFormula I and Formula IV are set forth in example below and generalizedin example-1 and example 2, respectively:

Example-1 Synthesis of Compound 1

A solution of metformin (free base obtained from HCl salt, 10 mmol)),((S)-2-ammonio-4-methylpentanoyl)-L-leucinate (10 mmol) andeicosapentaenoic acid (10 mmol)) in methanol (20 mL) was stirred at RTfor 5 hrs. The solvent was evaporated in vacuo and co-evaporated withTHF to dryness to afford compound in quantitative yield as a yellowsolid.

Example-2 Synthesis of Compound 3

To a solution of eicosapentaenoic acid (1 gms), BOP (1 gms) and DIPEA (1mL) in CH₂Cl₂ (40 mL) was added, L-Leucine methyl ester HCl (1 gms) andthe reaction mixture was stirred for overnight at RT. The reactionmixture was washed with water (40 mL) and the organic layer wasevaporated, crude was purified by column chromatography (20-40%EtOAc-hexanes) to afford intermediate ester (1 gms) as yellow syrup.

The above obtained ester was hydrolysed with NaOH (250 mg), in water (10mL) and THF (20 mL) at RT for overnight. The mixture was acidified with2N HCl (20 mL) and extracted with EtOAc (40 mL). The organic later wasdried over MgSO4, evaporated and purified by column chromatography toafford(S)-2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenamido)-4-methylpentanoicacid, compound 3 (1 gram) as yellow syrup.

Synthesis of compound 4

To a solution of compound 3 (10 mmol), BOP (10 mmol) and Et3N (13 mmol)in CH₂Cl₂ (20.0 mL) was added H₂N-Leu-OMe HCl (10 mmol) and stirred atRT for 12 hrs. The reaction mixture was diluted with water, organiclayer was separated and solvent was evaporated in vacuo. The crudeproduct was purified by flash chromatography (20-40% EtOAc/Hexanes)afforded EPA-Leu-Leu-OMe, compound 4 intermediate (6.5 mmol) as lightyellow syrup. The obtained intermediate was hydrolyzed with 1N NaOH andsubsequent quenching with dil HCl, and extracted with EtOAc (2×10 mL)provided((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenoyl)-L-leucyl-L-leucinate,compound 4 as yellow syrup.

Synthesis of Compound 5

Compound 4 (5 mmol) and metformin (5 mmol, free base) were mixed in THF(10 mL) and stirred at RT for 1 hr. The solution was evaporated in vacuoto dryness to afford compound 5 as a quasi solid in quantitative yield.

EQUIVALENTS

The present disclosure provides among other things compositions andmethods for treating diabetes and pre-diabetes and their complications.While specific embodiments of the subject disclosure have beendiscussed, the above specification is illustrative and not restrictive.Many variations of the systems and methods herein will become apparentto those skilled in the art upon review of this specification. The fullscope of the claimed systems and methods should be determined byreference to the claims, along with their full scope of equivalents, andthe specification, along with such variations.

INCORPORATION BY REFERENCE

All publications and patents mentioned herein, including those itemslisted above, are hereby incorporated by reference in their entirety asif each individual publication or patent was specifically andindividually indicated to be incorporated by reference. In case ofconflict, the present application, including any definitions herein,will control.

1. A compound of Formula I:

with at least one represented by X⁻, or a mixture thereof Wherein, X⁻independently represents


2. A compound of Formula II:

with at least one represented by X⁻, or a mixture thereof Wherein, X⁻independently represents


3. A compound of Formula III:

with at least one represented by X⁻, or a mixture thereof Wherein, X⁻independently represents


4. A pharmaceutical composition comprising a compound of claim 1 and apharmaceutically acceptable carrier, vehicle or diluent.
 5. Apharmaceutical composition comprising a compound of claim 2 and apharmaceutically acceptable carrier, vehicle or diluent.
 6. Apharmaceutical composition comprising a compound of claim 3 and apharmaceutically acceptable carrier, vehicle or diluent.
 7. Thepharmaceutical composition of claim 4, wherein said pharmaceuticalcomposition is formulated to treat an underlying etiology with aneffective amount administering to a patient in need by oraladministration, delayed release or sustained release, transmucosal,syrup, topical, parenteral administration, injection, subdermal, oralsolution, rectal administration, nanoparticle, buccal administration ortransdermal administration.
 8. The pharmaceutical composition of claim5, wherein said pharmaceutical composition is formulated to treat anunderlying etiology with an effective amount administering to a patientin need by oral administration, delayed release or sustained release,transmucosal, syrup, topical, parenteral administration, injection,subdermal, oral solution, rectal administration, nanoparticle, buccaladministration or transdermal administration.
 9. The pharmaceuticalcomposition of claim 6, wherein said pharmaceutical composition isformulated to treat an underlying etiology with an effective amountadministering to a patient in need by oral administration, delayedrelease or sustained release, transmucosal, syrup, topical, parenteraladministration, injection, subdermal, oral solution, rectaladministration, nanoparticle, buccal administration or transdermaladministration.
 10. The pharmaceutical composition of claim 7, whereinthe underlying etiology is diabetes mellitus, type 2 diabetes, type 1diabetes, obesity, lipid disorders, neuropathic pain,hypertriglyceridemia, NASH, NAFLD, hyperglycemia, pre-diabetes,dyslipidemia, hyperinsulinemia and insulin resistance.
 11. Thepharmaceutical composition of claim 8, wherein the underlying etiologyis diabetes mellitus, type 2 diabetes, type 1 diabetes, obesity, lipiddisorders, neuropathic pain, hypertriglyceridemia, NASH, NAFLD,hyperglycemia, pre-diabetes, dyslipidemia, hyperinsulinemia and insulinresistance.
 12. The pharmaceutical composition of claim 9, wherein theunderlying etiology is diabetes mellitus, type 2 diabetes, type 1diabetes, obesity, lipid disorders, neuropathic pain,hypertriglyceridemia, NASH, NAFLD, hyperglycemia, pre-diabetes,dyslipidemia, hyperinsulinemia and insulin resistance.